CZ200058A3 - Epothilone derivatives - Google Patents

Abstract

The present invention relates to compounds of the formula    Q is selected from the group consisting of    G is selected from the group consisting of alkyl, substituted alkvl, substituted or unsubstituted aryl, heterocyclo,    W is O or NR₁₅;
   X is O or H,H;
   Y is selected from the group consisting of O; H,OR₁₆; OR₁₇,OR₁₇; NOR₁₈; H,NOR₁₉; H,NR₂₀R₂₁; H,H; or CHR₂₂; OR₁₇ OR₁₇ can be a cyclic ketal;
   Z₁, and Z₂ are selected from the group consisting of CH₂, O, NR₂₃, S, or SO₂, wherein only one of Z and Z₂ is a heteroatom;
   B₁ and B₂ are selected from the group consisting of OR₂₄, or OCOR₂₅, or O₂CNR₂₆R₂₇; when B₁ is H and Y is OH, H they can form a six-membered ring ketal or acetal;
   D is selected from the group consisting of NR₂₈R₂₉, NR₃₀COR₃₁ or saturated heterocycle
   R₁, R₂, R₃, R₄, R₅, R₆, R₇, R₁₃, R₁₄, R₁₈, R₁₉, R₂₀, R₂₁, R₂₂, R₂₆, and R₂₇ are selected from the group H, alkyl, substituted alkyl, or aryl and when R₁ and R₂ are alkyl can be joined to form a cycloalkyl; R₃ and R₄ are alkyl can be joined to form a cycloalkyl;
   R₉, R₁₀, R₁₆, R₁₇, R₂₄, R₂₅, and R₃₁ are selected from the group H, alkyl, or substituted alkyl;
   R₈, R₁₁, R₁₂, R₂₈, R₃₀, R₃₂, R₃₃, and R₃₀ are selected from the group consisting of H, alkyl, substituted alkyl, aryl, substituted aryl, cycloalkyl, or heterocyclo;
   R₁₅, R₂₃ and R₂₉ are selected from the group consisting of H, alkyl, substituted alkyl, aryl, substituted aryl, cycloalkyl, heterocyclo, R₃₂C=O, R₃₃SO₂, hydroxy, O-alkyl or O-substituted alkyl, the pharmaceutically acceptable salts thereof and any hydrates, solvates or geometric, optical and stereoisomers thereof, with the proviso that compounds wherein
   W and X are both O; and
   R₁, R₂, R₇, are H; and
   R₃, R₄, R₆, are methyl; and
   R₈, is H or methyl; and
   Z₁, and Z₂, are CH₂; and
   G is 1-methyl-2-(substituted-4-thiazolyl)ethenyl; and
   Q is as defined above
are excluded.

Description

Epothilone derivatives

Technical field

The present invention relates to epothilone derivatives, processes for the preparation of these derivatives and intermediates for their preparation.

BACKGROUND OF THE INVENTION

Epothilones are macrolide compounds that find application in the pharmaceutical field. For example, epothilones A and B have been found to have the following structure:

Epothilone B R = Me exhibits a stabilizing effect on microtubules, similar to TAXOL, and therefore cytotoxic activity against rapidly malignant cells, such as tumor cells or other cellular hyperproliferative diseases, see Angew.

Chem. Int. Ed. Engl., 13/14,35 (1996).

SUMMARY OF THE INVENTION

The present invention relates to compounds of formula V

wherein Q is selected from the group consisting of

G is selected from the group consisting of alkyl, substituted alkyl, substituted or unsubstituted aryl, heterocyclo,

R-tt f 12 12

Ri f 12 12

R-12 f-12

oi ^r

R-I4

W is O or NR ₁₅ ;

X is an oxygen atom or Η, H;

Y is selected from the group consisting of oxygen; a hydrogen atom, OR = g; OR ₁₇ , OR ₁₇ ; NOR _lg ; a hydrogen atom, NOR ₁₉ ; a hydrogen atom, NR ₂ q R ₂₁ ; H, or CHR ₂₂ ; OR ₁₇ OR ₁₇ may be a cyclic ketal;

Z 1 and Z ₂ are selected from the group consisting of CH ₂ , oxygen atom, NR 23, sulfur atom or SO ₂ , with only one selected from the group consisting of CH ₂ , oxygen atom, NR 23, sulfur atom or SO ₂ ; ··

- Z 3 or Z ₂ may be a heteroatom;

B ₁ and B ₂ are selected from the group consisting of OR ₂₄ or OCOR ₂₅ or O ₂ CNR ₂₆ R ₂₇ if B - 1 is hydrogen and Y is OH, H may then form a six member ketal or acetal ring;

D is selected from the group consisting of NR ₂ gR ₂ g, _NR3 q COR ₃₁ or saturated heterocycle;

^R 1, ^R 2, ^R 3, ^R 4, ^R 5, ^R 6, ^R 7, ^R 13, ^R 14, ^R 18, ^R 19, ^R 20, ^R 21 ^R 22 ^R 26 ^{and R} 27 ^are selected from the group including hydrogen, alkyl, substituted alkyl or aryl, and when R 1 and R _{2 are} alkyl, they may be joined to form cycloalkyl; if R ₃ and R _{4 are} alkyl, they may be joined to form cycloalkyl;

R _9, R _10, R _g, R _17, R _24, R ₂₅ and R ₃₁ are selected from the group consisting of hydrogen, alkyl, substituted alkyl;

^R 8 ' ^R 11' ^R 12 ' ^R 28 ^{of R} 30' ^R 32 ' ^R 33 ^{and R} 30' ^are selected from the group consisting of hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, cycloalkyl or heterocyclo;

^R 15 ^R 23 and ^R 29 are selected from the group consisting of hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, cycloalkyl, heterocyclo, R ₃₂ C = O, R ₃₃ SO ₂ , hydroxy, O-alkyl or O-substituted alkoxy ;

and any salts, solvates and hydrates thereof.

Condition • ·

* This invention does not include compounds of formula V wherein

W and X are both oxygen atoms;

R _1, R ₂ and R ₇ are hydrogen atoms;

«

R ₃ , R ₄ and R 8 are methyl; and

R 8 is hydrogen or methyl; and

Z 1 and Z ₂ are CH ₂ ; and

G is 1-methyl-2- (substituted-4-thiazolyl) ethenyl;

Q has the same meaning as above.

DETAILED DESCRIPTION OF THE INVENTION

Listed below are definitions of various terms used to describe this invention. These definitions relate to terms as used throughout this disclosure, except as limited in particular cases, either individually or as part of a larger group.

The term alkyl refers to a linear or branched chain of unsubstituted hydrocarbon groups containing 1 to 20 carbon atoms, preferably 1 to 7 carbon atoms. The expression lower alkyl refers to unsubstituted C 1 -C 4 alkyl groups.

The term substituted alkyl refers to an alkyl group substituted with, for example, one to four substituents such as halogen, trifluoromethyl, trifluoromethoxy, hydroxy, alkoxy, cycloalkoxy, heterocycloxy, oxo, alkanoyl, aryloxy, alkanoyloxy, amino, alkylamino- , arylamino-, aralkylamino-, cycloalkylamino-, heterocycloamino, disubstituted amines in which the 2 amine substituents are selected from alkyl, aryl or arylalkyl, alkanoylamino-, aroylamino-, arylalkanoylamino, substituted alkanoylamino-alkyl, substituted-arylamino, substituted-arylamino, arylthio-, aralkylthio-, cycloalkylthio-, heterocyclothio-, alkylthiono-, arylthiono-, arylalkylthiono, alkylsulfonyl, arylsulfonyl, arylalkylsulfonyl, sulfonamido (e.g. SO ₂ NH ₂ ), substituted sulfonamino, nitro-, nitro- carboxy, carbamyl (e.g. CONH ₂ ), substituted with carbamyl (e.g. CONH-alkyl, CONH-aryl, CONH-arylalkyl or cases in which there are two substituents on the nitrogen atom selected from alkyl, aryl or aralkyl), alkoxycarbonyl, aryl, substituted aryl, guanidino and heterocyclo groups such as indolyl, imidazolyl, furyl, thienyl, thiazolyl, pyrrolidyl, pyridyl, pyrimidyl and the like. When it is mentioned above that the substituent is further substituted, it will be a halogen, alkyl, alkoxy, aryl or aralkyl substitution.

The term halogen refers to fluorine, chlorine, bromine and iodine.

The term aryl refers to monocyclic or bicyclic aromatic hydrocarbon groups having 6 to 12 carbon atoms in the ring moiety, such as phenyl, naphthyl, biphenyl and diphenyl groups, each of which may be substituted.

- 7 • ·

The term aralkyl refers to an aryl group on an alkyl group such as benzyl.

groups directly bound

The term substituted aryl refers to an aryl group substituted by, for example, one to four substituents such as alkyl, substituted alkyl, halogen, trifluoromethoxy, trifluoromethyl, hydroxy, alkoxy, cycloalkoxy, heterocyclooxy, alkanoyl, alkanoyloxy, amino, alkylamino, aralkylamino -, cycloalkylamino-, heterocycloamino-, dialkylamino-, alkanoylamino, thio-, alkylthio-, cycloalkylthio-, heterocyclothio-, ureido-, nitro-, cyano-, carboxy-, carboxyalkyl-, carbamyl-, alkoxycarbonyl-, alkylthiono-, sulfonyl-, thio-, thio-, thio- aryloxy and the like. The substituent may be further substituted by halogen, hydroxy, alkyl, alkoxy, aryl, substituted aryl, substituted alkyl or aralkyl.

that can be carbocyclic cyclobutyl, cyclooctyl,

The term cycloalkyl refers to optionally substituted, saturated hydrocarbon ring systems, preferably containing 1 to 3 rings and 3 to 7 ring carbon atoms, to be further condensed with an unsaturated ring containing 3 to 7 carbon atoms.

Exemplary groups include cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclodecyl, cyclododecyl and adamantyl. Exemplary substituents include one or more alkyl groups as listed above or one or more of the groups described above as alkyl substituents.

Terms heterocycle, heterocyclic and heterocycle • ·

10- refer to an optionally substituted, fully saturated or unsaturated, aromatic or non-aromatic ring group, which is, for example, a 4- to 7-membered bicyclic, 7- to

An 11-membered bicyclic or 10- to 15-membered tricyclic ring system having at least one heteroatom in at least one ring containing carbon atoms. Each ring of a heteroatom-containing heterocyclic group may have 1, 2 or 3 heteroatoms selected from nitrogen, oxygen and sulfur atoms, wherein the nitrogen and sulfur heteroatoms may be further oxidized and the nitrogen heteroatoms may be further quaternized. The heterocyclic group may be attached to any heteroatom or carbon atom.

Exemplary monocyclic heterocyclic groups include pyrrolidinyl, pyrrolyl, indolyl, pyrazolyl, oxetanyl, pyrazolinyl, imidazolyl, imidazolinyl, imidazolidinyl, oxazolyl, oxazolidinyl, isoxazolyl, thiazolyl, thiadiazolyl, thiazolidinyl, isothiazolinyl, isothiazolidinyl, isothiazolidinyl, isothiazolidinyl, isothiazolidinyl, isothiazolidinyl, isothiazolidinyl, , 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, 2-oxazepinyl, azepinyl, 4-piperidonyl, pyridyl, N-oxopyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, tetrahydropyranyl, tetrahydrothiopyranyl, tetrahydropyranylsulfone, morpholinyl, thiomorpholinyl, thiomorpholinyl, thiomorpholinyl, thiomorpholinyl 1,3-dioxolane and tetrahydro-1,1-dioxothienyl, dioxanyl, isothiazolidinyl, thietanyl, thiiranyl, triazinyl and triazolyl and the like.

Exemplary bicyclic heterocyclic groups include benzthiazolyl, benzoxazolyl, benzthienyl, quinuclidinyl, quinolinyl, quinolinyl-N-oxide, tetrahydroisoquinolinyl, isoquinolinyl, benzimidazolyl, benzpyranyl, indolizinyl, benzofuryl, chromonyl, coumarinyl, cinnolinyl, quinoxalinyl, quinoxalinyl

indazolyl, pyrrolopyridyl, furopyridinyl (such as furo [2,3-c] pyridyl, furo [3,1-b] pyridyl or furo [2,3-b] pyridyl), dihydroisoindolyl, dihydroquinazolinyl (such as 3,4-dihydro-4 -oxochinazolinyl), benzisothiazolyl, benzisoxazolyl, benzodiazinyl, benzofurazanyl, benzothiopyranyl, benzotriazolyl, benzotriazolyl, dihydrobenzofuryl, dihydrobenzothienyl, dihydrobenzothiopyranyl, dihydrobenzothiopyranylsulfon, dihydrobenzopyranyl, indolinyl, isochromanyl, isoindolinyl, naphthyridinyl, phthalazinyl, piperonyl, purinyl, pyridopyridyl, quinazolinyl, tetrahydroquinolinyl, thienofuryl, thienopyridyl, thienothienyl and the like.

Exemplary substituents include one or more alkyl groups as described above, or one or more groups described above as alkyl substituents. They also include smaller heterocycles such as epoxides and aziridines.

The term heteroatom includes oxygen, sulfur and nitrogen.

The compounds of formula V may form salts with alkali metals such as sodium, potassium and lithium, alkaline earth metals such as calcium or magnesium, with organic bases such as dicyclohexylamine, tributylamine, pyridine and amino acids such as arginine, lysine and the like. Such salts can be obtained, for example, by exchanging the carboxylic acid protons in the compounds of formula V when they contain the carboxylic acid with the desired ion in an environment in which the salt precipitates or in an aqueous environment followed by evaporation. Other salts may be formed as known to those skilled in the art.

The compounds of formula V form salts with various organic and inorganic acids. Such salts include those formed with hydrogen chloride, hydrogen bromide, methanesulfonic acid, hydroxyethane sulfonic acid, sulfuric acid, acetic acid, trifluoroacetic acid, maleic acid, benzenesulfonic acid, toluenesulfonic acid, and various other acids (eg, nitrates, phosphates, borates, tartarates) , citrates, succinates, benzoates, ascorbates, salicylates and the like). Such salts are formed by reacting a compound of formula V with an equivalent amount of acid in an environment in which the salt precipitates or in an aqueous medium and subsequent evaporation.

In addition, zwitterions (inner salts) are formed.

Compounds of formula V may also have prodrug forms. Any compound that is converted in vivo to provide a bioactive agent (i.e., a compound of Formula V) is a prodrug within the scope and spirit of the invention.

For example, compounds of Formula V may form an ester group of a carboxylate group. The carboxylate esters are conveniently formed by esterifying any of the carboxylic acid functionalities found on the disclosed ring structure (s).

Various forms of drug precursors are well known in the art. For examples of such precursor derivatives, see:

a) Design of Prodrugs, edited by H. Bundgaard, (Elsevier) (1985) and Methods in Enzymology, 42, 309-396, edited by K. Widder et al. (Academic Press) (1985);

(B) A Textbook of Drug Design and Development, edited by Krosgaard-Larsen and H. Bundgaard, Chapter 5, Design and Application of Prodrugs, H. Bundgaard, pp. 113-191 (1991);

c) H. Bundgaard, Advanced Drug Delivery Reviews, 8: 1-38 (1992);

d) H. Bundgaard et al., Journal of Pharmaceutical Sciences, 22, 285 (1998);

e) N. Kakeya et al., Chem. Phar. Bull., 32, 692 (1984).

It is further understood that solvates (eg, hydrates) of the compounds of Formula V are within the scope of this invention. Methods of solvation are generally known in the art.

Use and utility

Compounds of formula V are microtubule stabilizing agents. Thus, they can be used in the treatment of various cancers or other diseases associated with abnormal proliferation, including but not limited to:

cancers, including bladder, breast, colon, kidney, liver, lung, ovary, pancreas, stomach, cervix, thyroid and skin; including squamous cell carcinoma;

- hematopoietic tumors of lymphoid origin, including leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, ··· ··· ············ * »

Hodgkin's lymphoma, non-Hodgkin's lymphoma, hair cell lymphoma, and Burkett's lymphoma;

- hematopoietic tumors of myeloid origin, including acute and chronic myelogenous leukemia and promyelocytic leukemia;

tumors of mesenchymal origin, including connective tissue cancer and transverse striated muscle cancer;

- other tumors, including a pigment cell tumor, a testicular tumor, a tumor in immature tissues of various kinds, a germ cell-type cancer and a glioma;

- tumors of the central and peripheral nervous system, including nerve support tissue, germ-type glioma, glioma and schwannomas;

tumors of mesenchymal origin, including connective tissue cancer, transverse striated muscle cancer, and bone cancer;

- other tumors, including pigment cell tumor, xenoderma pigmentosum, keratoactanthoma, testicular tumor, thyroid cancer and immature tissue tumor of various kinds.

Compounds of formula V may also inhibit tumor angiogenesis, thereby affecting excessive malignant cell proliferation. These anti-angiogenic properties of the compounds of formula (V) may be useful in the treatment of certain forms of blindness associated with retinal vascular supply, arthritis, in particular. · Inflammatory arthritis, multiple sclerosis, restinosis and psoriasis.

The compounds of formula V can induce or suppress apoptosis, a physiological cell death process critical

Alternation of apoptotic pathways of disease changes in various general formulas (as limited to them), herpesvirus, smallpox, for routine development and homeostasis. contributes to the emergence and development of human disease. Apoptosis modulator compounds will be useful in the treatment of a variety of diseases in people with apoptosis abnormalities, including cancer (particularly follicular lymphomas, p53 mutations, breast, ovarian and ovarian cancer, hormone-dependent and ovarian cancer, such as familial adenomatous polyposis, but without viral infections (including

Epstein-Barr virus, Sindbis virus, and adenovirus, but not limited to, autoimmune diseases (including, but not limited to, systemic reddened caustic ulcer, immune-mediated glomerulonephritis, rheumatoid arthritis, psoriasis, inflammatory bowel disease and autoimmune diabetes) neurodegenerative diseases (including, but not limited to, Alzheimer's disease, AIDS-related dementia, Parkinson's disease, amyotropic lateral sclerosis, pigmentosa mucosal inflammation, spinal muscles, and cerebellum degeneration), AIDS, myelodysplastic syndromes, aplastic anemia, infarct anemia, infarct anemia with ischemic damage, stroke and reperfusion injury, arrhythmia, atherosclerosis, liver disease caused by toxin or alcohol, haematological disease (including, but not limited to, chronic anemia and aplastic anemia), muscle-degenerative disease (including, but not limited to, osteoporosis and arthritis), erní · erní erní erní erní erní erní erní erní erní erní erní erní

- 13 rhinosinusitis with aspirin sensitivity, cystic fibrosis, multiple sclerosis, kidney disease and cancer pain.

The compounds of the invention are also useful in combination with known anticancer and cytostatic agents and drugs, including radiation. When prepared in solid doses, such combination formulations utilize the compounds of this invention within the dosage range described below and other pharmaceutically active agents within the recommended dosage ranges. The compounds of formula (V) may be used sequentially with known anticancer and cytostatic agents and drugs, including radiation, if the combination composition is not suitable. Particularly effective are combinations of cytostatic drugs in which the second drug acts at a different stage of the cell cycle, e.g. in the S-phase, than the present compounds of formula V, which exert their effect in the G2-M phase.

E.g. thymidilate synthase inhibitors, DNA crosslinking agents, topoisomerase I and II inhibitors, DNA alkylating agents, ribonucleoside reductase inhibitors, cytotoxic factors such as TNF-α or growth factor inhibitors such as HER2 MAB receptors.

The present compounds may exist in multiple optical, geometric isomers and stereoisomers. All such isomers and mixtures thereof are within the scope of the invention.

The compounds of the invention may be processed.

With pharmaceutical excipients or solvents for oral, intravenous or subcutaneous administration. The pharmaceutical compositions may be formulated in conventional manner using solid or liquid excipients, diluents, and adjuvants suitable for the desired mode of administration. Orally, the compounds may be administered in the form of tablets, capsules, granules, powders, and the like. The compounds are administered in doses ranging from 0.05 to 200 mg.kg ^-1 per day, preferably less than 100 mg.kg ^-1 per day in a single dose or divided into 2 to 4 doses.

Preferred compounds

Particularly preferred compounds of formula V are those in which:

X is an oxygen atom,

Y is an oxygen atom,

Z 1 and Z ₂ are CH ₂ and W is NR ₁₅ .

Preparation method

Compounds of the general formula:

V are prepared according to

to

Scheme to

CHO

VI

OH O IX

Rs

OP, o X

³ K < ⁴

-f- “HOzC ^ YY ^ Rs

OP, O

XII

V (Q is an ethylene group) V (Q is an oxirane group), R ₄ , R ₅ , R 8, R 8 and R ₁₅ have the same meanings as above and wherein R 4 · ··

P ₁ is an oxygen protecting group.

Compounds of formula V wherein W is NR ₁₅ and X is an oxygen atom may be prepared according to Scheme 1. A compound of formula XII wherein Ρ _χ is an oxygen protecting group such as a target. butyldimethylsilyl may be prepared from a compound of formula VI by known methods (ie Nicolaou KC et al., Angew. Chem. Int. Ed. Engl., 36, 166-168 (1997)). The aldol reaction of a compound of formula XII and a compound of formula XIV provides a compound of formula XIII. The compound of formula XIV can be prepared by known methods (ie Schinzer D. et al., Eur. Chem. Chron., 1, 7-10 (1996)). The aldehyde of formula XVIII can be prepared from the compound of formula XV according to Scheme 1 or by known methods (ie Taylor RE et al., Tetrahedron Lett., 38, 2061-2064 (1997)). A compound of formula XIX can be prepared from a compound of formula XVIII by treatment with an amine using dehydration conditions such as a catalytic amount of p-toluenesulfonic acid and azeotropic removal of water. A compound of formula XX can be prepared from a compound of formula XIX by an allylating agent such as

A compound of formula XXI can be prepared from compounds of formulas XIII and XX by reaction with conventional amide bond binding agents (ie, DCC, BOP, EDC / HOBT, PyBrOP). A compound of formula XXII can be prepared from a compound of formula XXI by ring closure via either Grubbs (RuCl ₂ (= CHphenyl) (PCY ₃ ) ₂ ), see Grubbs RH et al., Angew. Chem. Int. 34, 2039 (1995) or a Schrock catalyst (see Schrock RR et al., J. Am. Chem. Soc., 112, 3875 (1990)) Removal of a protecting group in a compound of formula XXI using, for example, if Ρ _η processing with allylmagne with iumbromi dem.

target. butyldimethylsilyl, hydrogen fluoride in acetonitrile or tetra-n-butylammonium fluoride in THF provides a compound of formula V wherein Q is an ethylene group, W is NR ₁₅ , X is an oxygen atom and R ₃ , R ₄ , R _g and R _g have the same meaning as above. Regioselective epoxidation of a compound of formula V wherein Q is an ethylene group using dimethyldioxirane provides a compound of formula V wherein Q is an oxirane group, W is NR ₁₅ , X is an oxygen atom and R ₃ , R ₄ , R ₅ and R ₁₅ have the same meaning as above.

Scheme 2

Alternatively, a compound of formula VIII can be prepared by reacting a compound of formula XXIII with magnesium and an acid chloride (R RCt ^COCl) to give a compound of formula XXIV (see, for example, Heathcock C. et al., J. Org. Chem., 55. 1114-1177 (1990)) followed by ozonolysis to give a compound of formula VIII as shown in Scheme 2.

Scheme 3

XXVII

XXVIII

XIV • ·

The compound of formula XIV may optionally be prepared as depicted in Scheme 3. Reaction of a compound of formula XXV and pseudoephedrine provides a compound of formula XXVI. A compound of formula XXVII can be prepared from a compound of formula XXVI by alkylation with a pentenyl halide such as 5-bromopentene, according to the method of Meyers (ie, Meyers A. et al., J. Am. Chem. Soc., 116, 9361 and 9362 (1994)). A compound of formula XXVIII can be prepared from a compound of formula XXVII by reaction with a reducing agent such as lithium pyrrolidinylborohydride Oxidation of a compound of formula XXVIII using, for example, pyridinium chlorochromate provides a compound of formula XIV. an agent such as lithium triethoxyaluminium hydride.

Scheme 4

Alternatively, the compound of formula XX can be prepared from allylglicin as shown in Scheme 4. The nitrogen atom in allylglicin can be protected by methods known in the art to give a compound of formula XXIX, wherein P ₂ is

a suitable nitrogen protecting group, such as a tert. butoxycarbonyl. Alternatively, if R ₂ is not hydrogen, the compound of formula XXX can be prepared from a compound of formula XXIX by alkylation with an alkyl halide in the presence of a base such as sodium hydride. A compound of formula XXXI can be prepared from a compound of formula XXX using N, O-dimethylhydroxylamine and conventional coupling agents such as EDCI and HOBT. A compound of formula XXXII can be prepared from a hydroxamate of formula XXXI by treatment with an organometallic reagent such as an alkyl or aryl magnesium halide. Wittig olefinization of a compound of formula XXXII provides a compound of formula XXXIII (Wittig reagent is prepared as described by Danishefsky S. E et al., J. Org. Chem., 61, 7998-7999 (1996)). Removal of the nitrogen protecting group in the compound of Formula XXXIII by methods known in the art provides a compound of Formula XX.

Scheme 5 nr ₁₅ p ₂ a

RuCHžPOPte

Ri

NR ₁₅ P ₂

• · ϊ ϊ t t «« t t t t t t

A compound of formula V wherein W is NR ₁₅ , X is oxygen and G is a 1,2-disubstituted olefin can be prepared as depicted in Scheme 5. A compound of formula XXXV can be prepared by Wittig olefinization of a compound of formula XXXII. The compound of formula XXXIV can be prepared by methods known in the art. The compound of formula XXXVI can be prepared by removing the nitrogen protecting group in the compound of formula XXXV using methods known in the art. The compound of formula XXXVII can be prepared by the condensation reaction of a compound of formula XXXVI and a compound of formula XIII using conventional condensing agents such as EDCI and HOBT. The compound of formula XXXVIII may be of formula XXXVII by the methods of Scheme 1 for preparing a compound of formula XXII. Using the methods described in Scheme 1 (steps o and p), the compound of formula XXXVIII can be converted from the compound of the formula v to compounds of formula V wherein W is NR ₁₅ , X is oxygen and G is 1,2-disubstituted olefin.

is an atom

Scheme 6

XXXXII

V (Q is an ethylene group) in the (Q is an oxirane group _{in the} network

A compound of formula V wherein W and X are O and G is. The 1,2-disubstituted olefin can be prepared as shown in Scheme 6. A compound of formula XXXX can be prepared from a compound of formula XXXIX by treatment with an allylating agent such as allylmagnesium bromide. An enantiomerically pure compound of formula XXXX can be prepared using chiral reagents (see, for example, Taylor RE et al., Tetrahedron Lett. 38, 2061-2064 (1997); Nicolaou KC et al., Angew Chem. Int. Ed. Engl. 36, 166-168 (1997), Keck G. et al., J. Am. Chem. Soc., 115, 8467 (1993)). A compound of formula XXXXI can be prepared from compounds of formulas XXXX and XIII using known esterification methods such as DCC and DMAP. A compound of formula XXXXII can be prepared from a compound of formula XXXXI by a ring-closing olefin exchange reaction as described in Scheme 1 for the preparation of a compound of formula XXII. Compounds of formula V wherein W and X are oxygen atoms and G is a 1,2-disubstituted olefin can be prepared from a compound of formula XXXXII by deprotection (when Q is an ethylene group) and, if desired, epoxidation ( if Q is an oxirane group) as described above.

Scheme 7

A compound of formula V wherein W and X are oxygen atoms and G is alkyl, aryl, heteroaryl, bicycloaryl or bicycloheteroaryl can be prepared as depicted in Scheme 7. A compound of formula XXXXIV can be prepared by allylating a compound of formula XXXXIII wherein G is alkyl , substituted alkyl, aryl, heteroaryl, bicycloaryl or bicycloheteroaryl, by reaction with an allylating agent such as allyl magnesium bromide. A compound of formula XXXXV can be prepared from a compound of formula XXXXIV by esterification with a compound of formula XIII using, for example, DCC or DMAP. The compound of formula XXXXVI can be prepared from compound XXXXV by ring closure as described above. Following the methods described in Scheme 1 above, a compound of formula XXXXVI can be converted to a compound of formula V by deprotection followed by epoxidation.

Scheme 8

V ⁷ O a

XXXXIII

V ⁷

NRis

XXXXVII

V (Q is an ethylene group) V (Q is an oxirane group)

A compound of formula V wherein W is NR ₁₅ , X is oxygen and G is alkyl, aryl, heteroaryl, bicycloaryl or bicycloheteroaryl can be prepared as depicted in Scheme 8. A compound of formula XXXXVII can be prepared by reacting a compound of formula XXXXIII, wherein G is alkyl, substituted alkyl, aryl, heteroaryl, bicycloaryl or bicycloheteroaryl, and an amine under dehydration conditions. The compound of formula XXXXVIII can be prepared from compound XXXXVII by treatment with an allylating agent such as allyl magnesium bromide. A compound of formula XXXXIX can be prepared from a compound of formula XXXXVIII and a compound of formula XIII by conventional amide bond coupling methods using, for example, EDCI or HOBT. A compound of formula L can be prepared from a compound of formula XXXXIX by ring closure as described above. Following the methods described in Scheme 1 above, a compound of Formula L can be converted to compounds of Formula V by deprotection followed by epoxidation.

Scheme 9

A compound of formula V wherein X is O, W is NR _15, and G is

and D is selected from the group consisting of NR _2g R 29 ' ^NR 30 ^COR 31 and a saturated heterocycle (ie piperidinyl, morpholinyl, piperazinyl etc.) can be prepared as outlined in Scheme 9. A compound of formula LI can be prepared from a compound of formula XXXII by reductive amination using a primary or secondary amine and a reductive amine

agents such as sodium triacetoxyborohydride. Compounds of formulas LII, LIV and V can then be prepared by the following methods as described in Scheme 1 above.

Scheme 10

Alternatively, a compound of formula V wherein X is O is W is O or NR ₁₅ or oxygen and G is 12 12 and D is selected from the group consisting of NR 2 R 2 9/30 30 ^ 31 and saturated heterocycle (ie piperidinyl, morpholinyl , piperazinyl, etc.) can be prepared from a compound of formula V as shown in Scheme 10. A compound of formula V can be converted to a compound of formula LV by protecting the hydroxyl group with suitable protecting groups, such as a target. butyldimethylsilyl. A compound of formula LVI can be prepared from a compound of formula LV by ozonolysis. Treatment of a compound of the formula

LVI with an amine and a reducing agent such as sodium triacetoxyborohydride provides a compound of formula LVII. Removal of protecting groups from a compound of formula LVII, for example with hydrogen fluoride, provides a compound of formula V wherein X is an oxygen atom W is NR ₁₅ or an oxygen atom and

G is

Scheme 11

Compound of Formula V wherein W is NR ₁₅ X is O and G is

O

Ri3- _N Ay

R-14 can be prepared as outlined in Scheme 11. A compound of formula LVIII can be prepared from compounds of formula XXX by treatment with an amine and conventional amide bonding condensing agents such as EDCI and HOBT. A compound of formula LX can be prepared from a compound of formula LVIII by removal of a nitrogen protecting group using, for example, trifluoroacetic acid when P2 is a target. butoxycarbonyl group, followed by condensation of compounds of formulas LIX and XIII using conventional amide bond forming reagents such as EDCI and HOBT. The compound of formula LXI can be prepared from the compound of formula LX by ring closure. The compound of formula V can then be prepared from the compound of formula LXI by the methods described in Scheme 1 above.

Scheme 12

A compound of formula V wherein W is O,

X is an oxygen atom; and

G is 4 *

About _{13 η> Λ} ζ R ₁₄ may be prepared as shown in Scheme 12. A compound of formula LXII can be prepared from allylglycine by treatment with nitrous acid. A compound of formula LXIII can be prepared from a compound of formula LXII by treatment with an amine and conventional amide bonding condensing agents such as EDCI and HOBT. A compound of formula LXIV can be prepared from compounds of formulas LXIII and XIII using conventional amide bonding condensing agents such as EDCI and HOBT. The compound of formula LXV can be prepared from a compound of formula LXIV by ring closure. The compound of formula V can then be prepared from the compound of formula LXV by the methods described in Scheme 1.

ve j

·

Scheme 13

A compound of formula V wherein G is a 1,2-disubstituted ethyl group can be prepared from a compound of formula V wherein G is a 1,2-disubstituted ethylene group by hydrogenation with a catalyst such as palladium on carbon as shown in the scheme 13. In addition, a compound of formula V wherein G is a 1,2-disubstituted cyclopropyl group can be prepared from a compound of formula V wherein G is a 1,2-disubstituted ethylene group by cyclopropanation with diiodomethane and a zinc-copper pair, as described above. shows in scheme 4.

Scheme 14

Her

EtCfeC OH LXVI

H 3, C ^R 3kA ⁴

-VSr

OP, o XII

and

A compound of formula V wherein Z is an oxygen atom can be prepared as outlined in Scheme 14. A compound of formula LXVII can be prepared from an α-hydroxyester of formula LXVI and 3-buten-1-yltrifluoromethanesulfonate (or with 3-butenylbromide). and silver trifluoromethylsulfonate). The compound of formula LXVII can be reduced with a reducing agent such as diisobutylaluminum hydride to provide a compound of formula LXVIII. Alternatively, the compound of formula LXVIII can be obtained from the compound of formula LXVII by a two-step process comprising lithium borohydride reduction and pyridinium chlorochromate oxidation. This compound of formula LXVIII can be substituted with a compound of formula XIV of Scheme 1 to form a compound of formula LXIX. Further treatment of the compound of Formula LXIX as described above provides a compound of Formula V wherein Z 1 is an oxygen atom.

Scheme 15

χιι

Similarly, a compound of formula V wherein Z ₁ is NR _{23 can be} prepared as outlined in Scheme 15. A compound of formula LXXI can be prepared from the α-aminoester of formula LXX and 3-buten-1-yl bromide. The compound of formula LXXI can be reduced with a reducing agent such as diisobutylaluminum hydride to provide a compound of formula LXXII. Optionally, the compound of formula LXXII can be obtained from the compound of formula LXXI by a two-step process comprising lithium borohydride reduction and pyridinium chlorochromate oxidation. This compound of formula LXXII can be substituted with a compound of formula XIV of Scheme 1 to form a compound of formula LXXIII. Further treatment of a compound of Formula LXXIII as described above provides a compound of Formula V wherein Z 1 is NR ₂₂ .

• ·

Scheme 16

OH ^R p

XII

A compound of formula V wherein Z ₂ is an oxygen atom can be prepared as outlined in Scheme 16. A compound of formula LXXV can be prepared from a β-hydroxyester of formula LXXIV and an allylating agent such as allyl bromide (or allyl bromide and silver trifluoromethylsulfonate). The compound of Formula LXXV can be reduced with a reducing agent such as diisobutylammonium hydride to provide a compound of Formula LXXVI. Alternatively, the compound of formula LXXVI can be obtained from the compound of formula LXXV by a two step process comprising lithium borohydride reduction and pyridinium chlorochromate oxidation. This compound of formula LXXVI may be of formula XIV from Scheme 1 of formula LXXVII. Other compounds of Formula LXXVII as described above provide a compound of Formula V wherein Z ₂ is an oxygen atom.

substitute with compound to form compound processing • ·

Scheme 17

EtO _from C '

NHR

LXXVIII * 2323 * 88

LXXIX

OHCl 2 ξ * 23 ^R 3k, R4

HO ₂ C ^ Y ^ f ^ R ₅ OP, 0 XII

HO ₂ C

v <fJÍr?

OP, on LXXXI

Similarly, a compound of formula V wherein Z ₂ is NR ₂₂ can be prepared as outlined in Scheme 17. A compound of formula LXXIX can be prepared from a β-aminoester of formula LXXVIII and an allylating agent such as allyl bromide. The compound of formula LXXIX can be reduced with a reducing agent such as diisobutylaluminum hydride to provide a compound of formula LXXX. Alternatively, the compound of formula LXXX can be obtained from the compound of formula LXXIX by a two step process comprising lithium borohydride reduction and pyridinium chlorochromate oxidation. This compound of formula LXXX can be substituted with a compound of formula XIV of Scheme 1 to form a compound of formula LXXXI. Further treatment of the compound of Formula LXXXI as described above provides a compound of Formula V wherein Z ₂ is NR ₂₃ .

Scheme 18

A compound of formula V wherein W is oxygen or NR ₁₅ and Y is Η, H can be prepared as shown in Scheme 18. A compound of formula V can be converted to a compound of formula LXXXII wherein P ₄ and P ₅ are groups hydroxyl protecting groups, by treatment with an agent such as a target. butyldimethylsilyltrimethylsulfonate. The compound of formula LXXXIII can be prepared from a compound of formula by treatment with Lawesson's reagent. A compound of formula LXXXIV can be prepared from a compound of formula LXXXIII using a reducing agent such as tri-n-butyltin hydride when W is oxygen, or by treatment with methyl iodide and sodium borohydride when W is NR ₁₅ . Removal of protecting groups from a compound of formula LXXXIV using, for example, hydrogen fluoride when P ₄ and P _{5 are} silyl groups, provides a compound of formula V wherein W is O or NR ₁₅ and Y is H, H.

• w • 4 * 4

Scheme 19

The compound of formula (V) wherein W and Y are oxygen atoms and is alkyl or substituted alkyl may be an alkyl or substituted alkyl. The compound of formula V can be protected to give a compound of formula LXXXV wherein P ₅ and P _g are hydroxyl protecting groups by treatment with a reagent such as a target. butyldimethylsilyltrifluoromethanesulfonate. A compound of formula LXXXVI can be prepared from a compound of formula LXXXV by treatment with a reducing agent such as sodium borohydride. A compound of formula LXXXVII can be prepared from a compound of formula LXXXVI by introducing a hydroxyl protecting group wherein P ₇ is, for example, p-methoxybenzyl, using p-methoxybenzyltrichloroacetimidate. Removal of the protecting groups P ₅ and Pg from the compound of formula LXXXXVII, using, for example, hydrogen fluoride in pyridine, when P ₅ and Pg are targets. butyldimethylsilyl groups, provides a compound of formula LXXXXVIII, which can then be selectively protected using, for example, a target. butyldimethylsilyl chloride, to form a compound of formula LXXXXIX, wherein Ρθ is the tert. butyldimethylsilyl group. A compound of formula C can be prepared from a compound of formula LXXXXIX by treatment with a base such as lithium diisopropylamide and subsequent treatment with an alkylating agent such as methyl iodide. The compound of formula (C) may be protected to form a compound of formula (Cl) wherein P 1 is a hydroxyl protecting group by treatment with a reagent such as a target. butyldimethylsilyltrifluoromethanesulfonate. A compound of formula C11 can be prepared from a compound of formula Cl, by removing the P ₇ group using, for example, DDQ when P _{7 is a} p-methoxybenzyl group. A compound of formula V wherein W and Y are oxygen atoms and is alkyl or substituted alkyl may be prepared from a compound of formula C11 by oxidation using, for example, TPAP / NMO followed by deprotection, using for example

hydrogen fluoride when Pg and Pg are silyl groups. This compound of formula V can be further oxidized with dimethyldioxiran according to Scheme 1 to provide the corresponding epoxy compound of formula V.

Scheme 20

V 'is an ethylene group)

A compound of formula V wherein X is an oxygen atom and Q is an olefin can be prepared from a compound of formula V wherein X is an oxygen atom and Q is an oxirane ring by treatment with a reactive metallocene such as titanocene, zirconocene or niobocene as described in Scheme 20 (see, for example, R. Schobert and U. Hohlein, Synlett, 465-466 (1990)).

Scheme 21

O OH O V (W is an oxygen atom)

OH

A compound of formula V wherein X is an oxygen atom and W is NR 1 - g, wherein R 1 is a hydrogen atom can be prepared from a compound of formula V wherein both X and W are oxygen atoms as shown in Scheme 21. A compound of formula (III) can be prepared from a compound of formula (V) wherein both X and W are oxygen atoms by forming a pi-allyl palladium complex using, for example, palladium tetrakistriphenylphosphine and subsequent treatment with sodium azide (see, for example, Murahashi S.-I. et al. J. Org. Chem., 54, 3292 (1989)). Subsequent reduction of a compound of formula (III) with a reducing agent such as triphenylphosphine provides a compound of formula (CIV). A compound of formula V wherein X is oxygen and W is NR ₁₅ wherein R ₁₅ is hydrogen can be prepared from a compound of formula CIV by macrolactamization using, for example, diphenylphosphoryl azide or bromotripyrrolidinophosphonium hexafluorophosphate (PyBroP).

Scheme 22

(W is an oxygen atom)

(W is NR ₁₅ )

A compound of formula V wherein X is O and W is NR ₁₅ wherein R ₁₅ is alkyl, substituted alkyl, aryl, heteroaryl, cycloalkyl, heterocyclyl, O-alkyl, O-substituted alkyl may be prepared from a compound of formula V, wherein both X and W are oxygen atoms as depicted in Scheme 22. A compound of formula CV can be prepared from a compound of formula V wherein both X and W are oxygen atoms by forming a pi-allyl palladium using for example palladiumtetrakisphenylphosphine by treatment with a primary amine . A compound of formula V wherein X is O and W is NR ₁₅ , wherein R ₁₅ is alkyl, substituted alkyl, aryl, heteroaryl, cycloalkyl, heterocyclyl, hydroxyl, O-alkyl,

O-substituted alkyl, can be prepared from a compound of formula V by macrolactamization using, for example, a complex followed by and subsequent

9

9

A diphenylphosphoryl azide or bromotripyrrolidinophosphonium hexafluorophosphate (PyBroP). In the case where R ₁₅ is a hydroxyl group, it may be necessary to remove a protecting group such as a target. butyldimethylsilyl, from an intermediate wherein R ₁₅ is O-tert. butyldimethylsilyl.

In vitro monitoring of the biological activity of compounds of Formula V was performed as follows:

In vitro polymerization of tubulin

Double-cycled calf tubulin was prepared according to the Williams and Lee method (see Williams RC Jr. and Lee JC, Preparation in Tubulin from Methods. Methods in Enzymology, 85, part D: 376-385 (1982)) and before use. stored in liquid nitrogen. Quantification of tubulin polymerization efficiency was achieved by the following modified procedure of Swindell et al. (See Swindell CS, Krauss NE, Horwitz SB and Ringel I., Biologically active taxol analogues with deleted A-ring side chain substituents and variable C-2 'configurations. J. Med. Chem., 34, 1176-1184 (1991) ). These adjustments partially resulted in the expression of tubulin polymerization efficiency as an effective concentration for any given compound. In this method, various concentrations of compound in polymerization buffer (0.1 M MES, 1 mM EGTA, 0.5 mM MgCl ₂ , pH 6) are added to tubulin at 37 ° C in a Beckman Model DU 7400 micro-cuvette cell. , 6). A final microtubular protein concentration of 1.0 mg.ml ^-1 and a compound concentration of usually 2.5, 5.0 and 10 μΜ are used. The default ODD, measured every 10 seconds, was calculated by a program embedded in the instrument after manually determining the start and end times of the linear region including the.

at least 3 time points. Under these conditions, the linear deviations were usually <10 ^-6 , the slopes ranged from 0.03 to 0.002 absorbance units per minute and the highest absorbance was 0.15 absorbance units. The effective concentration (ECq q1) is determined as an interpolated concentration capable of producing a slope of 0.01 OD.min ^-1 and is calculated according to the equation: EC _{0 01} = concentration / slope. ECq ₀₁ values are expressed as the mean of the standard deviation obtained from 3 different concentrations. The ECq ₀₁ values of the compounds of the invention fall within the range of 0.01 to 1000 μΜ.

Cytotoxicity (in vitro)

Cytotoxicity was determined on human colon carcinoma HCT-116 cells by MTS (3- (4,5-dimethylthiazol-2-yl) -5- (3-carboxymethoxyphenyl) -2- (4-sulfenyl) -2H-tetrazolium, inert salt) as reported by TL Riss et al. in Comparison of MTT, XTT, and a novel tetrazolium compound MTS for in vitro proliferation and chemosensitivity assays, Mol. Biol. Cell, 3., (Suppl.), 184a (1992). Cells were seeded at 400.0 cells per well in 96-well microfiltration plates and after 24 hours drug was added and gradually diluted. The cells were incubated at 37 ° C for 72 hours, at which time they were stained with tetrazolium, MTS was added at a concentration of 333 µg.ml ^-1 (final concentration), in combination with the electron condensation agent phenazine methosulfate at 25 µΜ (final concentration). ). Enzyme dehydrogenase in living cells reduces MTS to a form that absorbs light at 4900 nm, which can be determined spectrophotometrically. The greater the absorbance, the greater the number of viable cells. The results were expressed as IC ₅₀ , which is the concentration of drug required to prevent cellular malignancy (ie absorbance at 450 nm) in 50% of the cells.

compared to untreated control cells. IC ₅₀ values for compounds of the invention fall within the range of 0.01 to 1000 nM.

The following examples illustrate the invention.

DETAILED DESCRIPTION OF THE INVENTION

Example 1

[4S- [4R *, 7S *, 8R *, 9R *, 15R * (E)]] - 4,8-Dihydroxy-5,5,7,9-tetramethyl-16- [1-methyl-2- ( 2-methyl-4-thiazolyl) ethenyl] -1-aza-13 (E) -cyclohexadecene-2,6-dione

A. N - [(2-Methyl) -1-propenyl] morpholine

To stirred morpholine (165.5 g, 1.9 mol) was added isobutyraldehyde (173 mL, 1.9 mol) at a rate that did not allow the reaction mixture to exceed 30 ° C. After complete addition, the reaction mixture was stirred at room temperature for 2 hours, then the flask was fitted with a Dean-Stark trap and heated to 160 ° C for 20 hours. The reaction mixture was then cooled to room temperature and the flask was fitted with a vigreux column distillation apparatus. Distillation under high vacuum gave 135 g (50%) of Compound A as a clear colorless oil. MS (+ + H, 142).

B. 2,2-Dimethyl-3-oxopentanal

To a stirred solution of propionyl chloride (44 mL, 0.50 mol) in ether (135 mL) at 0 ° C under nitrogen was added a solution of Compound A (69 g, 0.50 mol) in ether over 45 min. The reaction mixture was filtered (50 mL). Volatiles 2 ⁽ (135 mL). After complete addition with condenser for 2 hours and room for 16 hours, and filter cake washed with ether was distilled off in vacuo. The residue (80 mL) and the solution was adjusted to pH 4. Ether was added. The reaction mixture was poured into a separatory funnel, the layers were separated, and the aqueous layer was extracted with ether (5x100 mL) .The combined organic phases were dried (MgSO 4), filtered and evaporated in vacuo. distilled under high vacuum to give 10.4 g (16%) of Compound B as a clear colorless oil MS (MH, 127).

dissolved in H _by 0

C. 4-tert. Butyldimethylsilyloxy-5,5-dimethyl-6-oxo-1-octene

To (25.7 g, atmosphere of a solution of (-) - β-methoxydiisopinocamphenylborane mmol) in ether (80 mL) at 0 ° C under nitrogen was added a 1.0 M solution of allyl magnesium bromide in ether (77 mL, 77) at 0 ° C over 1.5 h. mmol). The reaction mixture was stirred at 25 ° C for 1 hour and then evaporated in vacuo. The residue (2x150 mL) and extracts were under Celite. The combined extracts were afforded B-allyldiisopropinocamphenylborane. This material was dissolved in ether (200 mL) and cooled under nitrogen to extract with pentane nitrogen filtered through then evaporated in vacuo

-100 ° C. A solution of compound B (11.42 g, 89 mmol) in ether (90 mmol) was then added over 1 h at -78 ° C. The reaction mixture was stirred for an additional 0.5 hours and then methanol (1.5 mL) was added. The reaction mixture was brought to room temperature, treated with 3N NaOH solution (32 mL) and 30% H 2 O 2 (64 mL) and then held at reflux for 2 hours. The reaction mixture was cooled to room temperature, the layers were separated, and the organic phase was washed with H ₂ O (500 mL). The combined aqueous washes were re-extracted with ether (2 x 100 mL). The combined organic extracts were washed with saturated aqueous NaCl (100 mL), dried (MgSO _4), filtered and evaporated under vacuum. The residue was dissolved in CH 2 Cl 2 (250 mL), cooled to 0 ° C, and diisopropylethylamine (93 mL, 535 mmol) was added. The target solution is then slowly poured into the stirred solution. butyldimethylsilyltrifluoromethanesulfonate

260 mmol) such that the temperature did not rise above 10 ° C. After the addition, the reaction mixture was poured into H ₂ O (650 mL), the layers were separated and the aqueous layer was washed with ΟΗ ₂ Ο _{1 2} (2x650 mL). The combined organics were dried (Na ₂ SO ₄ ), filtered and evaporated in vacuo. The residue was purified by flash chromatography eluting with hexanes followed by 10% EtOAc / hexanes to give 17.2 g (78%) of Compound C as a clear colorless oil. An enantiomeric difference of 94% was found by ¹ H NMR analysis of the Mosher alcohol ester. ¹³ C NMR (CDCl ₃ , 80 MHz) d 215.8, 136.1, 116.5, 52.8, 39.0, 31.9, 26.0, 22.4, 20.1, 18.1 , 7.6, -3.6, -4.4.

added (69 g, complete

D. 3-tert. Butyldimethylsiloxy-4 _/ 4-dimethyl-5-oxoheptanal

A solution of Compound C (10.8 g, 38.0 mmol) in CH 2 Cl 2 was bubbled with _{3 3} at -78 ° C until the solution was permanently blue (1 hour). O ₂ was then bubbled through for 15 minutes, followed by N ₂ bubbled for 30 minutes, after which time

the solution became clear. Then triphenylphosphine (10 g, 38 mmol) was added and the reaction mixture was warmed to -35 ° C and allowed to stand for 16 hours. The volatiles were evaporated in vacuo and the residue purified by flash chromatography eluting with 8% EtOAc / hexanes to afford 8.9 g (74%) of Compound D as a clear colorless oil. ¹ H NMR (CDCl ₃ , 300 MHz) d 9.75 (m, 1H), 4.53 (t, J = 4.8 Hz, 1H),

3.40-3.60 (m, 4H), 1.10 (s, 3H), 1.07 (s, 3H), 0.98 (t,

J = 7.0 Hz, 3H), 0.83 (s, 9H), 0.07 (s, 3H), 0.04 (s, 3H).

E. 3-tartaric acid. butyldimethylsiloxy-4,4-dimethyl-5-oxoheptane

To a solution of compound D (3.90 g, 13.6 mmol) in tert. butanol (75 mL) was added 2-methyl-2-butene (5.85 mL, 55.2 mmol) and then a solution of sodium chlorite (4.61 g, 40.8 mmol) and monobasic was added dropwise at room temperature. sodium phosphate (2.81 g, 20.4 mmol) in H ₂ O (15 mL). The reaction mixture was stirred for 0.5 hour and then the solvents were removed in vacuo. H ₂ O (150 mL) was added to the residue followed by extraction with EtOAc (3x150 mL). The combined organic extracts were dried (MgSO ₄ ), filtered and the volatiles removed in vacuo. The residue was purified by flash chromatography (20%).

EtOAc / hexanes / 1% AcOH to give 3.79 g (92%) of Compound E as a clear colorless viscous oil. IMS (.delta. + H, 303).

F. (R, R) -N- (2-Hydroxy-1-methyl-2-phenethyl) -N, 2- (S) -dimethyl-6-heptenamide

A suspension of LiCl (6.9 g, 0.16 mol) and preformed lithium diisopropylamide (Aldrich, 2.0 M solution in hepta46 / ethylbenzene / THF, 27.6 mL, 55 mmol) in additional THF (70 mL) was added at - 78 ° C was treated dropwise with a solution of (R, R) -N- (2-hydroxy-1-methyl-2-phenethyl) -N-methylpropionamide (6.0 g, 27 mmol, Meyers AG et al., J. Am Chem Soc., 116, 9361 (1994)) in THF (30 mL) for 10 minutes. The light yellow reaction mixture was stirred at -78 ° C (1 hour), at 0 ° C (15 minutes) and at 25 ° C (5 minutes) before being re-cooled to 0 ° C and treated with a solution of 5- bromo-1-pentene (4.8 mL, 40 mmol) in THF (5 mL). The reaction mixture was stirred at 0 ° C (24 h), poured into a saturated aqueous solution of NH 4 Cl (100 mL) and EtOAc (100 mL). The two phases were separated and the aqueous phase was further extracted with EtOAc (3 x 100 mL). The organic extracts were combined, washed with saturated aqueous NaCl (200 mL), dried (Na ₂ SO ₄ ), and evaporated in vacuo. Flash chromatography (SiO ₂ , 4.0x25 cm, 2% MeOH-CHCl-g) gave compound F (6.9 g, 88%) as a yellow oil. MS (ESI ⁺ ): 290 (M + H) ^&lt; ^{+ &gt};, MS (ESI ^- ): 288.2 ([delta] - H).

G. (S) -2-Methyl-6-heptenol

A 250 mL round bottom flask was sequentially charged with pyrrolidine (2.6 mL, 30 mmol) and BH ₃ -THF complex (1.0 M in THF, 31 mL, 30 mmol) at 0 ° C. The borane-pyrrolidine complex was heated to 25 ° C (1 hour), cooled to 0 ° C and treated dropwise with n-butyllithium (1.0 M in hexane, 19 mL, 30 mmol) over 30 minutes while carefully maintained an internal temperature of 5.5 ° C. The reaction mixture was stirred at 0 ° C for an additional 30 minutes before a solution of compound F (3.0 g, 10 mmol) in THF (23 mL) was added dropwise over 10 minutes. The reaction mixture was stirred at 25 ° C (6 hours) before the reaction was quenched by dropwise addition of aqueous 3N HCl solution (25 mL). The reaction mixture was then poured over an aqueous 1 N HCl solution (200 mL) and extracted with Et 2 O

(4x80 ml). The combined organics were washed with a 1: 1 mixture of saturated aqueous NaCl and 1N aqueous HCl (2 x 150 mL) and evaporated in vacuo. Aqueous NaOH (1N, 200 mL) was added to the residue and the suspension was stirred for 30 minutes. The mixture was extracted with Et ₂ O (3x100 mL) and the combined ether layers were washed with a 1: 1 mixture of saturated aqueous NaCl and 1 N NaOH (2x200 mL), dried (Na ₂ SO ₄ ) and evaporated in vacuo. Flash chromatography (SiO ₂ , 4.0x25 cm, gradient elution with 15-25% Et ₂ O-pentane solution) gave compound G (1.26 g, 95%) as a colorless oil, [α] ²⁵ D -H ( c 12, ch ₂ or ₂ ).

H. (S) -2-Methyl-6-heptenal

A solution of compound G (0.24 g, 1.9 mmol) in CH ₂ Cl ₂ (6 mL) was treated with pyridinium chlorochromate (0.61 g, 2.8 mmol) and the reaction mixture was stirred at 25 ° C for 5 min. hours. The resulting dark brown viscous slurry was passed through a silica gel-Celite column (Celite 1.0 x 1 cm at the top of SiCl 2).

I, 0x5 cm, eluting with 50 mL of CH ₂ Cl ₂ ). The solvent was evaporated in vacuo to give crude compound H (0.15 g, 63%) as a colorless oil which was sufficiently pure to be used in subsequent reactions. ¹ H NMR (300 MHz, CD ₂ Cl ₂ ): d 9.62 (s, 1H), 5.88-5.68 (m, 1H), 5.13-4.92 (m, 2H), 2 37-2.24 (m, 1H), 2.15-2.05 (m, 2H), 1.62-1.78 (tn, 1H), 1.51-1.32 (m, 3H) 1.07 (d, 3H, J = 7.0 Hz).

I. (3S, 6R, 7S, 8S) -3-tert. Butyldimethylsiloxy-4,4,6,8-tetramethyl-7-hydroxy-5-oxo-12-tridecene

To a previously prepared solution of LDA (Aldrich, 2.0 M solution in heptane / ethylbenzene / THF, 3.8 mL, 7.6 mmol) in additional THF (25 mL) at -78 ° C was added dropwise during the A solution of compound E (1.0 G, 3.4 mmol) in THF (5 mL) was added over 1 min. The reaction mixture was stirred at -78 ° C (10 minutes), warmed to -40 ° C (20 minutes), and re-cooled to -78 ° C before compound H (0.56 g, 4) was added. , 4 mmol) in THF (5 mL). The reaction mixture was warmed to -40 ° C, stirred for 1 hour and diluted with NH ₄ Cl (50 mL). They were extracted with EtOAc (4x50 mL), with a saturated aqueous solution of two layers, and the aqueous phase was separated

The combined organic layers were washed with saturated aqueous NaCl solution (100 mL), dried (Na ₂ SO ₄ ) and evaporated in vacuo. Flash chromatography (SiO ₂ , 2.5x20 cm, gradient elution 2 to 5% MeOH-CHCl 3) followed by HPLC separation (YMC S-10, ODS, 30x500 mm column, eluting with MeOH at 20 mL.min ^-1 ) gave the desired syn-aldol product, compound and undesired diastereoisomer (0.32 g, compound E (about 10%)).

I (0.60 g, 43%) 22%) along with the starting material

MS (ESI ⁺ ): 879.3 (2 M + Na);

MS (ESI) 427.3 (M-H) -.

451.2 (M + Na) ^&lt; ^{+ &gt};, 429.2 (M + H)

The stereochemistry was confirmed by direct comparison of ¹ H NMR and ¹ H NMR of the following ester derivative (used in the synthesis of epothilone C) with the same intermediate previously described by KC Nicolaou et al. in Angew. Chem. Int. Ed. Engl., 36, 166 (1997).

J. (S) -2- [N- [tec. butyloxy) carbonyl] amino] -4-pentene

A solution of L-2-amino-4-pentenoic acid (NovaBiochem, 3.0 g, 26 mmol) in THF-H ₂ O (1: 1, 200 mL) was treated sequentially with NaHCO 3 (6.6) at 0 ° C. g, 78 mmol) and di-tert. butyldicarbonate (10.4 g, 1.8 mmol). The reaction mixture was warmed to 25 ° C and stirred for 16 hours. The pH of the mixture was adjusted to * ·

pH 4 by carefully adding a saturated aqueous citric acid solution at 0 ° C and the mixture was extracted with EtOAc (4x50 mL). The combined organic layers were washed with saturated aqueous NaCl solution (75 mL), dried (Na ₂ SO ₄ ) and evaporated in vacuo. Flash chromatography (SiO ₂ , 4.0x6 cm, gradient elution with 5-10% MeOH-CHCl 3) gave Compound J (5.5 g, 99%) as a colorless oil. NS (ESI ^- ): 429.3 (2M-H), 214.1 (MH).

K. (S) -2- [N ² - [(tert -butyloxy) carbonyl amino-N-methoxy-N-methyl-4-pentenamide

A solution of compound J (2.9 g, 13 mmol) in CHCl 3 (55 mL) at 0 ° C was treated sequentially with N, O-dimethylhydroxylamine hydrochloride (1.4 g, 15 mmol), 1-hydroxybenzotriazole (2.0 g, 15 mmol), 4-methylmorpholine (4.4 mL, 40 mmol) and 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (3.4 g, 18 mmol). The reaction mixture was slowly warmed to 25 ° C, stirred for 16 hours, and diluted with H ₂ O (100 mL). The two layers were separated and the aqueous phase was extracted with EtOAc (3x75 mL). The combined organic phases were washed with 5% aqueous HCl (100 mL), saturated aqueous NaHCO ₃ (100 mL), saturated aqueous NaCl (100 mL), dried (Na ₂ SO ₄ ) and evaporated in vacuo. Flash chromatography (SiO ₂ , 3.0x20 cm, gradient elution with 25 to 50% EtOAc-hexane) gave compound K (2.5 g, 71%) as a colorless oil. MS (ESI ⁺ ): 258.9 (M + H) ⁺ , 202.9 (M-isobutylene), 158.9 (M-BOC), MS (ESI ^- ): 257.2 (MH) ^- .

L. (S) -3- [N - ((tert-Butyloxy) carbonyl] amino] -5-hexen-2-one

A solution of compound K (2.5 g, 1.0 mmol) in THF (65 mL) at 0 ° C was treated with methylmagnesium bromide (3.0 M solution).

in Et ₂ O, 8.1 mL, 2.4 mmol). The reaction mixture was stirred at 0 ° C (2.5 hours) and carefully poured into saturated aqueous NH ₄ Cl solution (100 mL). The two layers were separated and the aqueous phase was extracted with EtOAc (3x75 mL). The combined organic extracts were washed with saturated aqueous NH ₄ Cl (75 mL), H ₂ O (75 mL), saturated aqueous NaCl (75 mL), dried (MgSO ₄ ) and evaporated in vacuo. Flash chromatography (SiO ₂ , 3.0x20 cm, gradient elution with 10-25% EtOAc-hexane) gave (S) -2- [N - [(tert-butyloxy) carbonyl] amino] -5-hexen-2-one ( 2.2 g, 67%) as a colorless oil. MS (ESI ⁺ ): 213.9 (M + H) ⁺ , 157.9 (M-isobutylene), 113.9 (M-BOC), MS (ESI ^- ): 212.2 (MH).

M. (S) -4- [3- [N - [(tert-Butyloxy) carbonyl] amino] -2-methyl-1 (E), 5-hexadienyl] -2-methylthiazole

A solution of 2-methyl-4-thiazolylmethyldiphenylphosphine oxide (2.5 g, 8.0 mmol, Danishefsky et al., J. Org. Chem., 61, 7998 (1996)) in THF (38 mL) at -78 ° C treated dropwise over 5 minutes with n-butyllithium (1.6M solution in hexane). The resulting bright orange mixture was stirred at -78 ° C and treated with a solution of 3.8 mmol) in THF (5 mL). After the cooling bath was removed

5.2 mL, 8.4 mmol). for 15 minutes of compound 10 minutes

L (0.81 g, at

-78 ° C and the reaction mixture was allowed to warm to 25 ° C (2 hours). The mixture was poured into saturated aqueous NH ₄ Cl solution (50 mL) and the two layers were separated. The aqueous phase was extracted with Et ₂ O (3x50 mL) and the combined organic extracts were washed successively with H ₂ O (75 mL), saturated aqueous NaHCO 3 (75 mL), saturated aqueous NaCl (75 mL), dried (Na ₂ SO ₄ ) and evaporated in vacuo. Flash chromatography (SiO ₂ , 4.0x30 cm, gradient elution with 10-20% EtOAc-hexane) gave compound M (0.23 g, 18%) as a colorless oil along with the recovered starting ketone (20-30%). ). MS (ESI ⁺ ): 309.1 (M + H) ⁺ , 253.0 (M-isobutylene), MS (ESI -): 307.3 (MH).

N. (S) -4- (3-Amino-2-methyl-1 (E), 5-hexadienyl) -2-methylthiazole

Compound M (0.15 g, 0.49 mmol) was treated with 4.0 N HCl in 1,4-dioxane (5 mL) at 0 ° C under argon. The volatiles were evaporated in vacuo and the resulting white foam was dissolved in cold saturated NaHCO ₃ (3 mL). This solution was extracted with EtOAc (4 x 10 mL) and the combined EtOAc portions were dried (Na ₂ SO ₄ ) and evaporated in vacuo. Flash chromatography (SiO ₂ , 1.0x5 cm, 5 to 10% MeOH-CHCl ₃ gradient) gave compound N (88 mg, 88%) as a colorless oil. MS (ESI ⁺ ): 209.0 (M + H) ^&lt; ^{+ &gt};, MS (ESI): 207.2 (MH) &lt; - &gt;.

O. (3S, 6R, 7S, 8S) -N- (S) - [1- (2-Methyl-4-thiazolyl) -2-methyl-1 (E) -5-hexadien-3-yl] -3 -target. Butyldimethylsiloxy-4,4,6,8-tetra-methyl-7-hydroxy-5-oxo-12-tridecenamide

A solution of compound M (88 mg, 0.42 mmol) in DMF (1.3 mL) at 0 ° C was treated sequentially with compound I (0.15 g, 0.35 mmol), 1-hydroxybenzotriazole (49 mg, 0). , 36 mmol) and 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (0.10 g, 0.52 mmol). The reaction mixture was slowly warmed to 25 ° C, stirred for 15 hours, and diluted with H ₂ O (3 mL). The mixture was extracted with EtOAc (3 x 10 mL) and the combined organic phases were washed with 5% aqueous HCl (10 mL), saturated aqueous NaHCO ₃ (10 mL), saturated aqueous NaCl (10 mL), dried (Na ₂ SO _4). ) and evaporated in vacuo. Flash chromatography (SiO ₂ , 1.5x20 cm, 2.5% mixture) · ·

MeOH-CHCl ₃ ) afforded compound 0 (0.17 g, 77%) as a white foam. MS (ESI ⁺ ), 619.3 (M + H) &lt; + & gt ^; .

P. [4S- [4R *, 7S *, 8R *, 9R *, 15R * (E)]] - 4-tert. Butyldimethylsiloxy-8-hydroxy-5,5,7,9-tetramethyl-16- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -1-aza-13 (E) -cyclohexadecen-2, 6-dione

A solution of compound O (17 mg, 27 mmol) in degassed benzene (8.0 mL) was treated with Grubb's catalyst [bis (tricyclohexylphosphine) benzylidene ruthenium dichloride, Strem Chemicals, 11 mg, 14 mmol) under argon. The reaction mixture was stirred at 25 ° C for 15 hours and re-treated with additional catalyst (5.0 mg, 4.5 mmol). After a further 7 hours, the benzene was evaporated in vacuo and the black viscous residue was passed through a pad of silica gel (1.0x3 cm) eluting with Et ₂ O (25 mL). The eluent was evaporated in vacuo to give a divisible 5: 1 (E / Z) geometric isomer mixture. PTLC (SiO ₂ , 1 mm plate, 2 eluting with 1: 1: 1 hexane-toluene-ethyl acetate) gave the E-isomer of compound P (5.1 mg, 34%) and the corresponding Z-isomer (1.0 g, 6 , 7%). For compound P: MS (ESI ⁺ ): 1181.7 (2M + H) ⁺ , 591.4 (M + H) ⁺ . For

Z-isomer: MS (ESI ⁺ ): 1181.5 (2M + H) ⁺ , 613.2 (2M + Na) ⁺ , 591.2 (M + H) ⁺ , MS (ESI ^- ): 589.3 ( MH).

Q. [4S- [4R *, 7S *, 8R *, 9R *, 15R * (E)]] - 4,8-Dihydroxy-5,5,7,9-tetramethyl-16- [1-methyl-2] - (2-Methyl-4-thiazolyl) ethenyl] -1-aza-13 (E) -cyclohexadecene-2,6-dione

To a vial of compound P (2.3 g, 0 ° C) was added an acid content of 3.55 cm 3 filled with a solution of 3.9 mmol) in CH ₂ Cl ₂ (0.4 mL) at trifluoroacetic acid (0.1 mL). The reaction mixture was sealed under argon and stirred at 0 ° C. After hours, the volatiles were removed at 0 ° C under a constant flow of argon. To the residue was added saturated aqueous NaHCO 3 (1 mL) and EtOAC (1 mL), and the two layers were separated. The aqueous phase was extracted with EtOAc (4 x 1 mL) and the combined ethyl acetate layers were dried (Na ₂ SO ₄ ) and evaporated in vacuo. PTLC (SiO ₂ , 20x10x0.025 cm, eluting with 5% MeOH-CHCl ₃ ) gave [4S- [4R *, 7S *, 8R *, 9R *, 15R * (E)]] - 4,8-dihydroxy- 5,5,7,9-tetra-methyl-16- [1-methyl-2- (2-methyl-4-thiazolyl) -ethenyl] -1-aza-13 (E) -cyclohexadecene-2,6-dione (1.3 mg, 68%) as a white film. MS (ESI): 953.5 (2M + H) ⁺ , 477.3 (M + H) ⁺ , MS (ESI -): 475.5 (MH).

Example 2

The following compounds can be prepared according to the previously described reaction schemes:

[1S- [1R *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] - 7,11-dihydroxy-8,8,10,12,16-pentamethyl-3- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -4,13,17-trioxabicyclo [14.1.0] heptadecane-5,9-dione, [1S- [1R *, 3R *] (E), 7R *, 10S *, 11R *, 12R *, 16S *]] - 7,11-dihydroxy-8,8,10,12-tetramethyl-3- [1-methyl-2- (2-methyl)] -4-thiazolyl) ethenyl] -4,13,17-trioxabicyclo [14.1.0] heptadecane-5,9-dione, [4S- [4R *, 7S *, 8R *, 9R *, 15R * (E ]]] - 4,8-Dihydroxy-5,5,7,9,13-pentamethyl-16- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -1,10-dioxa-13 -cyclohexadecene-2,6-dione, [4S- [4R *, 7S *, 8R *, 9R *, 15R * (E)]] - 4,8-dihydroxy-5,5,7,9-tetramethyl-16 - [1-methyl-2- (2-methyl-4-thiazolyl) 54-ethenyl] -1,10-dioxa-13-cyclohexadecene-2,6-dione, [1S- [IR], 3R * (E ), 7R *, 10S *, 11R *, 12R *, 16S *]] -7,11-dihydroxy-8,8,10,12,16-pentamethyl-3- [1-methyl-2- (2-methyl-4) -thiazolyl) ethenyl] -4,14,17-trioxabicyclo [14.1.0] heptadecane-5,9-dione, [1S- [IR], 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] - 7,11-dihydroxy-8,8,10,12-tetramethyl-3- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -4,14,17-trioxabicyclo [14,1, O] heptadecane-5,9-dione, [4S- [4R *, 7S *, 8R *, 9R *, 15R * (E)]] - 4,8-dihydroxy-5,5,7,9,13- pentamethyl-16- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -1,11-dioxa-13-cyclohexadecene-2,6-dione, [4S- [4R *, 7S *, 8R] *, 9R *, 15R * (E)]] - 4,8-Dihydroxy-5,5,7,9-tetramethyl-16- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] - 1,11-dioxa-13-cyclohexadecene-2,6-dione, [1S- [IR *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] - 7,11-dihydroxy- 8,8,10,12,16-pentamethyl-3- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -4,17-dioxabicyclo [14.1.0] heptadecane-5.9 -dione, [1S- [IR *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] - 7,11-dihydroxy-8,8,10,12-tetramethyl-3 - [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -4,17-dioxabicyclo [14.1.0] heptadecane-5,9-dione, [1S- [1R *, 3R * (E)] , 7R *, 10S *, 11R *, 12R *, 16S *]] - 7,11-dihydroxy-3,8,8,10,12,16-hexamethyl-3- [1-methyl-2- (2-met-55)

hyl-4-thiazolyl) ethenyl] -4,17-dioxabicyclo [14.1.0] heptadecane-5,9-dione, [1S- [1R *, 3R * (E), 7R *, 10S *, 11R *] 12R *, 16S *]] - 7,11-dihydroxy-3,8,8,10,12-pentamethyl-3- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -4,17- dioxabicyclo [14.1.0] heptadepan-5,9-dione, [4S- [4R *, 7S *, 8R *, 9R *, 15R * (E)]] - 4,8-dihydroxy-5, 5,7,9,13,16-hexamethyl-16- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -1-oxa-13-cyclohexadecene-2,6-dione, [4S- [4R *, 7S *, 8R *, 9R *, 15R * (E)]] - 4,8-dihydroxy-5,5,7,9,16-pentamethyl-16- [1-methyl-2- (2 -methyl-4-thiazolyl) ethenyl] -1-oxa-13-cyclohexadecene-2,6-dione, [1S- [IR], 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] -7,11-dihydroxy-6,8,8,10,12,16-hexamethyl-3- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -4,17-dioxabicyclo [14.0] , 1,0] heptadecane-5,9-dione, [1S- [1R *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] -7,11-dihydroxy-6,8 8,10,12-pentamethyl-3- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -4,17-dioxabicyclo [14.1.0] heptadecane-5,9-dione, [4S- [4R *, 7S *, 8R *, 9R *, 15R * (E)]] - 4,8-dihydroxy-5,5,7,9-tetramethyl-16- [1-methyl-2- ( 2-methyl-4-thiazolyl) ethenyl] -1-aza-13-cyclohexadecene-2,6-dione, [1S- [1R *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] - 7,11-dihydroxy -4,8,8,10,12,16-hexamethyl-3- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -4-aza-17-oxabicyclo [14.1.0] 56 • heptadecan-5,9-dione, [1S- [1R *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *] ]] -7,11-dihydroxy-4,8,8,10,12-pentamethyl-3- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -4-aza-17-oxabicyclo 1,0] heptadecane-5,9-dione, [4S- [4R *, 7S *, 8R *, 9R *, 15R * (E)]] - 4,8-dihydroxy-1,5,5,7, 9,13-hexamethyl-16- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -1-aza-13-cyclohexadecene-2,6-dione, [4S- [4R *, 7S *] , 8R *, 9R *, 15R * (E)]] - 4,8-Dihydroxy-1,5,5,7,9-pentamethyl-16- [1-methyl-2- (2-methyl-4-thiazolyl)] (ethenyl) -1-aza-13-cyclohexadecene-2,6-dione, [1S- [1R *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] - 7, 11-dihydroxy-8,8,10,12,16-pentamethyl-3- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -13-aza-4,17-dioxabicyclo [14,1,0 Heptadecan-5,9-dione, [1S- [IR *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] -7,11-dihydroxy-8,8,10,12 -tetramethyl- 3- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -13-aza-4,17-dioxabicyclo [14.1.0] heptadecane-5,9-dione, [4S- [4R] *, 7S *, 8R *, 9R *, 15R * (E)]] - 4,8-dihydroxy-5,5,7,9,13-pentamethyl-16- [1-methyl-2- (2-methyl)] -4-thiazolyl) ethenyl] -10-aza-1-oxa-13-cyclohexadecene-2,6-dione, [4S- [4R *, 7S *, 8R *, 9R *, 15R * (E)]] - 4,8-dihydroxy-5,5,7,9-tetramethyl-16- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -10-aza-1-oxa-13-cyclohexadecene-2 , 6-dion, [1S- [1R *, 3R * (E), 7R *, 10S *, 11R *] 12R *, 16S *]] -7,11-dihydroxy-8,8,10,12,16-pentamethyl-3- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -14-aza- 4,17-dioxabicyclo [14.1.0] heptadecane-5,9-dione, [1S- [1R *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] - 7,11-dihydroxy-8,8,10,12-tetramethyl-3- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -14-aza-4,17-dioxabicyclo [14,1,0 heptadecan-5,9-dione, [4S- [4R *, 7S *, 8R *, 9R *, 15R * (E)]] - 4,8-dihydroxy-5,5,7,9,13-pentamethyl -16- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -1-aza-1-oxa-13-cyclohexadecene-2,6-dione, [4S- [ 4R *, 7S *, 8R *, 9R *, 15R * (E)]] - 4,8-dihydroxy-5,5,7,9-tetramethyl-16- [1-methyl-2- (2-methyl- 4-thiazolyl) ethenyl] -1-aza-1-oxa-13-cyclohexadecene-2,6-dione, [1S- [IR], 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] -Nphenyl-7,11-dihydroxy-8,8,10,12,16-pentamethyl-5,9-dioxo4,17-dioxabicyclo [14.1.0] heptadecane-3-carboxamide, [1S- [IR *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] - NF-phenyl-7,11-dihydroxy-8,8,10,12-tetramethyl-5,9-dioxo4 17-dioxabicyclo [14.1.0] heptadecane-3-carboxamide, [4S- [4R *, 7S *, 8R *, 9R *, 15R * (E)]] - N-Phenyl-4,8-dihydroxy -5,5,7,9,13-pentamethyl-2,6-dioxo-1-oxa-13-cyclohexadecene-16-carboxamide, [4S- [4R *, 7S *, 8R *, 9R *, 15R * ( E)]] - N-Phenyl-4,8-dihydroxy-5,5,7,9-tetramethyl-2,6-dioxo-1-oxa-13-cyclohexadecene-16-carboxamide,

i [1 S - [IR *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] -7.11 *

Dihydroxy-8,8,10,12,16-pentamethyl-3- [1-methyl-2- (2-methyl-4-thiazolyl) cyclopropyl] -4,17-dioxabicyclo [14,1,0] heptadecane- 5,9-dione and [1S- [1R *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] - 7,11-Dihydroxy-8,8,10,12-tetramethyl- 3- [1-methyl-2- (2-methyl-4-thiazolyl) cyclopropyl] -4,17-dioxabicyclo [14.1.0] heptadecane-5,9-dione.

Example 3

[1S- [1R *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] - 7,11-Dihydroxy-8,8,10,12,16-pentamethyl-3- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -4-aza-17-oxabicyclo [14.1.0] heptadecane-5,9-dione.

A. (3S, 6R, 7S, 8S, 12R, 13S, 15S) -15-azido-12,13-epoxy-4,4,6,8,12,16-hexamethyl-7-hydroxy-17- ( 2-methyl-4-thiazolyl) -5-oxo-16-heptadecene.

A solution of epothilone B (0.35 g, 0.69 mmol) in degassed

THF (4.5 mL) was treated with a catalytic amount (80 mg, mmol) of tetrakis (triphenylphosphine) palladium (0) and the suspension was stirred under argon at 25 ° C for 30 minutes. The resulting bright yellow homogeneous solution was all worked up at once

with a solution of sodium azide (54 mg, 0.83 mmol) in degassed H ₂ O (2.2 mL). The reaction mixture was heated to 45 ° C for 1 hour, diluted with H ₂ O (5 mL) and extracted with EtOAc (4x75 mL). The organic extracts were washed with saturated aqueous NaCl solution (15 mL), dried (Na ₂ SO ₄ ) and evaporated in vacuo. The residue was purified by flash chromatography (SiO ₂ , 3.0x15 cm, 95: 5.0: 0.5 CHCl ₃ -MeOH-AcOH) to give compound A (0.23 g, 61%) as a colorless oil. MS (ESI ^&lt; ^{+ &} gt ^; ): 551 (M + H) &lt; + & gt ^; , MS (ESI ^&lt; ^- & gt ^; ): 549 (MH).

B. (3S, 6R, 7S, 8S, 12R, 13S, 15S) -15-Amino-12,13-epoxy-4,4,6,8,12,16-hexamethyl-7-hydroxy-17- ( 2-methyl-4-thiazolyl) -5-oxo-16-hepadiene

A solution of compound A (0.23 g, 0.42 mmol) in THF (4.0 mL) at 25 ° C was treated with H ₂ O (23 mL, 1.25 mmol) and polymer-supported triphenylphosphine (Aldrich, polystyrene). cross-linked 2% DVB, 0.28 g, 0.84 mmol). The resulting suspension was stirred at 25 ° C under an argon atmosphere (32 hours), filtered through Celite and evaporated in vacuo. The residue was purified by flash chromatography, gradient elution from 95: 5.0: 0.5

CHCl ₃ -MeOH-AcOH) to give compound B (96 mg, 44%) as a colorless oil. MS (ESI ⁺ ): 525.2 (M + H) + ¹ ',

MS (ESI -): 523.4 (M-H).

1.5x10 cm, 90: 10: 1.0 (SiO ₂ , do

Alternatively, absolute EtOH was added to a 25 mL round bottom flask charged with Compound A (0.26 g, 0.47 mmol) and PtO ₂ (0.13 g, 50% by weight) under argon. The resulting black mixture was stirred under a H ₂ atmosphere for 10 minutes, after which time the system was purged with N ₂ and another portion of PtO ₂ (65 mg, 25% by weight) was added. Once again, the reaction mixture was stirred under a H ₂ atmosphere for 10 hours. System se

Then purged with N ₂ and the reaction mixture was filtered through a pad of Celite eluting with CH ₂ Cl ₂ (3x25 mL). The solvents were evaporated in vacuo and the residue was purified as above to give compound B (0.19 g, 75%).

Alternatively, a solution of compound A (20 mg, 36 mmol) in THF (0.4 mL) under argon was treated with triphenylphosphine (19 mg, 73 mmol). The reaction mixture was heated to 45 ° C, stirred for 14 hours and cooled to 25 ° C. The resulting iminophosphorane was treated with ammonium hydroxide (28%, 0.1 mL) and the reaction mixture was heated once more to 45 ° C. After 4 hours, the volatiles were removed in vacuo and the residue was purified as above to give compound B (13 mg, 70%).

C. [1S- [1R *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] -7,11-Dihydroxy-8,8,10,12,16-pentamethyl-3- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -4-aza-17-oxabicyclo [14.1.0] heptadecane-5,9-dione

A solution of compound B (0.33 g, 0.63 mmol) in degassed DMF (250 mL) at 0 ° C under argon was treated with solid NaHCO ₃ (0.42 g, 5.0 mmol) and diphenylphosphoryl azide (0.15 g, 5.0 mmol). 54 mL, 2.5 mmol). The resulting suspension was stirred at 4 ° C for 24 hours, diluted at 0 ° C with phosphate buffer (250 mL, pH 7) and extracted with EtOAc (5x100 mL). The organic extracts were washed with 10% aqueous LiCl solution (2 x 125 mL), dried (Na ₂ SO ₄ ) and evaporated in vacuo. The residue was first purified by flash chromatography (SiO ₂ , 2.0x10 cm, gradient elution 2 to 5% MeOH-CHCl 3) and then purified using Chromatotron (2 mm SiO ₂ , GF rotor, gradient elution 2 to 5%). MeOH-CHCl3) to give the title compound (0.13 g, 40%) as a colorless oil. ¹ H NMR (CDCl ₃ , 400 MHz): δ 6.98 (s, 1H), 6.71 (d, 1H, NH, J = 8.1 Hz),

6.56 (s, 1 H), 4.69 to 4.62 (m, 1H), 4.18 to 4.12 (m, 1H), 4.01 to 3.96 (m, 1H), 3.86 (s, 1H), 3.38 to 3.34 (m, 1H), 2.82 (dd, 1 H, J = 5.6, 6.0 Hz), 2.71 (s, 3H), 2.58 (with, 1H), 2.43 (dd, h, J = 9.0, 14.5 Hz), 3.34 (dd F 1H, J = 3.0, 14.5 Hz), 2.14 (s, 3H), 2.05-1.92 (m, 2H) 1.82 up to 1.41 (series multiplets, 7H), 1.35 (s, 3H), 1,28 (with, 3H), 1.18 (d, 3H, J = 6.8 Hz) 1.14 (s, 3H). 1.00 (d, 3H, J = 6.8 Hz); MS (ESI ⁺ ): 507.2 (M + H) ^&lt; ^{+ &gt};; MS (ESI ^-): 505.4 (M- • H) -.

Example 4

A method for reducing the oxirane ring of epothilone and epothilone analogs

Chopped chips were placed in a two-necked flask and vacuum dried under argon. (250 mg, 1.0 mmol) was added as an almost aliquot (3.5 mL, magnesium (24 mg, 1.0 mmol) was consumed.) Flask by flame and cooled under bis (cyclopentadienyl) titanium dichloride followed by anhydrous THF ( The stirred suspension was degassed by deep vacuum and the flask was refilled with argon.The red suspension became dark, turning to a uniformly dark green after 1.5 hours with all the magnesium metal removed.

0.70 mmol, 3.5 eq) and cooled to -78 ° C under argon. After this solution, epothilone A (99 mg, 0.20 mmol, 1.0 eq) was added. The reaction mixture was warmed to room temperature and stirred for 15 minutes. The volatiles were evaporated in vacuo and the residue chromatographed twice on silica (25 g) using 35% EtOAc / hexanes as the carrier liquid to give 76 mg (80%) of epothilone C as a light yellow viscous oil.

Example 5

[1S- [1R *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] - 7,11-Dihydroxy-8,8,10,12-tetramethyl-3- methyl-2- (2-methyl-4-thiazolyl) ethenyl] -4-aza-17-oxabicyclo [14.1.0] heptadecane-5,9-dione

A. (3S, 6R, 7S, 8S, 12R, 13S, 15S) -15-Azido-3,7-dihydroxy-12,13-epoxy-4,4,6,8,16-pentamethyl-17- (2- methyl-4-thiazolyl) -5-oxo-16 (E) -heptadecene

To a solution of epothilone A (4.97 g, 10.1 mmol, 1.0 eq) in degassed THF (100 mL) at room temperature was added tetrakis (triphenylphosphine) palladium (0) (1.17 g, 1.01) mmol, 0.10 eq) and stirred for 30 minutes under argon. To this reaction mixture was added sodium azide (0.980 g, 15.1 mmol, 1.5 eq) followed by degassed water (10 mL). The reaction mixture was heated at 45 ° C for 1 hour, cooled to room temperature, diluted with ethyl acetate (300 mL), and further diluted with water (150 mL). The aqueous layers were extracted with ethyl acetate (3x100 mL). The combined organic extracts were washed with brine (150 mL), dried (sodium sulfate), filtered and evaporated in vacuo. The oily residue was purified by flash chromatography on silica gel (eluting with 0 to 5% methanol / chloroform with 0.1% acetic acid) to afford Compound A (1.84 g, 34.0% yield) as a glassy solid. MS (ESI ⁺ ): 537 (M + H) ^&lt; ^{+ &gt};, MS (ESI ^- ): 535 (MH) ^&lt; ^{- &gt};.

B. (3S, 6R, 7S, 8S, 12R, 13S, 15S) -15-Amino-3,7-dihydroxy-12,13-epoxy-4,4,6,8,16-pentamethyl-17- (2- methyl-4-thiazolyl) -5-oxo-16 (E) -heptadecene

To a solution of compound A (1.85 g, 3.44 mmol, 1.0 eq) in absolute ethanol (137 mL) was added platinum oxide (0.980 g, 4.30 mmol, 1.25 eq). The reaction mixture was vigorously stirred at room temperature under a hydrogen balloon for 16 hours. The reaction mixture was filtered and the filtrate was evaporated in vacuo. The oily residue was purified by preparative HPLC (YMC S-15 ODS 50x500 mm column, 45 minutes / gradient, 0-100% B, 50 ml min ^-1, retention time 17 minutes, A = 0.1% acetic acid / 5 % acetonitrile / 95% water, B = 0.1% acetic acid / 5% water / 95% acetonitrile). The appropriate fractions were evaporated in vacuo and the residue was lyophilized from aqueous acetonitrile to give compound B (1.33 g, 76.0% yield) as a colorless solid.

MS (ESI ^&lt; ^{+ &} gt ^; ): 511 (M + H) ^&lt; ^{+ &gt};, MS (ESI ^&lt; ^- & gt ^; ): 509 (MH) ^- .

C. [1S- [1R *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] -7,11-dihydroxy-8,8,10,12-tetramethyl-3- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -4-aza-17-oxabicyclo [14.1.0] heptadecane-5,9-dione

Compound B (0.860 g, 1.68 mmol, 1.0 eq) was dissolved in anhydrous DMF (0.00250 M, 672 mL) and degassed for 1 hour at room temperature. The solution was cooled to 0 ° C and anhydrous sodium bicarbonate (1.13 g, 13.4 mmol, 4.0 eq) and diphenylphosphoryl azide (1.85 g, 6.72 mmol, 8.0 eq) were added under an argon atmosphere. .). The reaction mixture was kept under argon at 4 ° C and stirred for 16 hours. Then, the reaction mixture was cooled to -60 ° C and pH 7 phosphate buffer (400 mL) was added slowly to quench the reaction. The temperature was kept below -30 ° C. The mixture was allowed to warm slowly to room temperature and extracted with ethyl acetate (1 L). The aqueous layer was washed with ethyl acetate (4x300 mL). The organic extracts were combined, washed with 10% LiCl solution (500 mL), dried (sodium sulfate), filtered and evaporated in vacuo. The oily residue was purified by preparative HPLC (YMC S-15 ODS 50x500 mm column, 45 min / gradient elution, 0 to 100% B, 50 mL min ^-1 , retention time = 35 min, A = 5% acetonitrile / 95% water B = 5% water / 95% acetonitrile). The appropriate fractions were evaporated in vacuo and the residue was lyophilized from aqueous acetonitrile to give the title compound (0.220 g, 26.0% yield) as a colorless solid.

MS (ESI ⁺ ): 493 (M + H) ^&lt; ^{+ &gt};; MS (ESI ^- ): 491 (MH) ^{& lt} ; - ^&gt;.

Example 6

Me

[4S- [4R *, 7S *, 8R *, 9R *, 15R * (E)]] - 4,8-Dihydroxy-5,5,7,9,13-pentamethyl-16- [1-methyl-2] - (2-methyl-4-thiazolyl) ethenyl] -1-aza-13 (Z) -cyclohexadecene-2,6-dione

Tungsten chloride (0.19 g, 0.49 mmol, 0.5 eq) was dissolved in THF (5.0 mL) and the solution was cooled to -78 ° C. A solution of n-butyllithium in hexane (1.6 M, tungsten reagent at room temperature (0.79 mL, 0.079 mmol) was added in one portion, and reaction minutes (2.0 mL) were added to the compound.

0.63 mL, 1.0 mmol, 1.0 eq) and allowed to warm to room temperature over 20 minutes (the solution became dark green while warming to room temperature). 0.1 M solution thus prepared

2.0 mmol) was added

4C (0.020 g, 0.039 mmol, 1.0 eq). the mixture was stirred at room temperature for a period of time and then quenched with saturated NaHCO ₃ solution. This solution was diluted with water (10 mL) and the solution was extracted with CH ₂ Cl ₂ (4x20 mL). The combined organic extracts were dried (Na ₂ SO ₄ ), filtered and evaporated in vacuo. The inorganics were removed by passing the residue through a pad of silica gel (eluting with 19/1 CHCl3 / MeOH). The eluent was evaporated in vacuo. The residue was purified by PHPLC (YMC S-5 ODS, 30-100% B, A = 5% aqueous CH? CN, B = 95% aqueous CH? CN, 3 ml min ^-1, 220 nm, 30 minutes; gradient elution) and the appropriate fractions were evaporated in vacuo. The sticky solid was lyophilized from aqueous acetonitrile to give the title compound (4.3 mg, 29%) as a white solid. TLC: Rf = 0.57 (9/1 CHCl3 / MeOH, visible by UV light), HRMS: (M + H) 4-, calc 491.29436, found 491.2934.

Example 7

• [• S - [1R *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] - 7,11-Dihydroxy-8,8,10,12,16-pentamethyl-3 - [1-methyl-2- (2-hydroxymethyl-4-thiazolyl) ethenyl] -4-asa-17-oxabicyclo [14.1.0] heptadecane-5,9-dione

A. N-Oxide [1S-11R *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] -7,11-dihydroxy-4,8,8,10,12- pentamethyl-3- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -4-asa-17-oxabicyclo [14.1.0] heptadecane-5,9-dione

A solution of epothilone B (2.0 g, 3.9 mmol) in CH ₂ Cl ₂ (30 mL) was treated with 3-chloroperoxybenzoic acid (1.0 g, 5.9 mmol) at 25 ° C for 2 h under argon. ). An additional 0.5 g (3.0 mmol) of 3-chloroperoxybenzoic acid was added and the reaction mixture was stirred for 2 hours. The reaction mixture was filtered and the filtrate was evaporated in vacuo. The residue was dissolved in EtOAc (100 mL), washed with saturated aqueous solution of NaHCOg (75 mL), 5% aq Na ₂ SOg (75 mL), H ₂ O (75 mL), dried (Na ₂ SO ₄₎ and evaporated under vacuum. The residue was purified by flash chromatography (SiO ₂ , 4.5x30 cm, gradient elution with 2 to 10% MeOH-CHCl-g solution) to give compound A (1.04 g, 50%) as a white solid. MS (ESI ⁺ ), 524.3 (M + H) ^&lt; ^{+ &gt};; MS (ESI ^- ): 522.5 (MH).

B. [1S- [1R *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] -7,11-Dihydroxy-8,8,10,12,16-pentamethyl- 3- [1-methyl-2- (2-hydroxymethyl-4-thiazolyl) ethenyl] -4,17-dioxabicyclo [14.1.0] heptadecane-5,9-dione [epothilone F]

To a solution of compound A (0.46 g, 0.88 mmol) in CH ₂ C1 ₂ (10 mL) in a resealable tube under Argon was added 2,6-lutidine (0.82 mL, 7.0 mmol) and trifluoroacetic anhydride (0.87 mL, 6.2 mmol). The reaction vessel was sealed under argon, heated to 75 ° C (12 minutes), cooled to 25 ° C, and the volatiles were removed under a constant stream of N ₂ . The reaction tube was then connected to the high vacuum pump for 15 minutes. The resulting residue was dissolved in MeOH (10 mL) and treated with ammonium hydroxide (28-30% NH ₄ in H ₂ O, 1 mL). The mixture was heated to 45 ° C (10 minutes) and the volatiles were evaporated in vacuo. The crude reaction mixture was purified by HPLC (YNC S-15 ODS 30x500 mm column, 50% acetonitrile-water solution, isocratic conditions, flow rate 20 ml / min retention time 28 min). The appropriate fractions were evaporated in vacuo and the residue was lyophilized from aqueous acetonitrile to give compound B (0.22 g, 48%) as a white solid.

MS (ESI ⁺ ): 524.3 (M + H) ⁺ , 1047.6 (2M + H) ^&lt; ^{+ &gt};, (MH) &^lt; ^{+ &gt};.

white solids. MS (ESI): 522.5

C. (3S, 6R, 7S, 8S, 12R, 13S, 15S) -15-Azido-3,7-dihydroxy-12,13-epoxy-4,4,6,8,12,16-hexamethyl-17- ( 2-hydroxymethyl-4-thiazolyl) -5-oxo-16 (E) -heptadecene

A solution of compound B (0.18 g, 0.34 mmol) in degassed THF (3.0 mL) was treated with a catalytic amount (40 mg, 3.4.10 ^-2 mmol) of tetrakis (triphenylphosphine) palladium (0) and the mixture was treated with was stirred under argon at 25 ° C for 30 minutes. The resulting bright yellow homogeneous solution was all treated all at once with an azide solution

0.41 mmol) in degassed H ₂ O (1.5 mmol) was heated at 45 ° C for 1 hour, diluted with H ₂ O (5 mL) and extracted with EtOAc (4x10 mL). The organic extracts were washed with a saturated aqueous NaCl solution (15 mL), dried (Na ₂ SO ₄ ) and evaporated in vacuo. The residue was purified by flash chromatography (SiO ₂ , 2.5x15 cm, gradient elution 95: 5.0: 0.5 CHCl 3 · -MeOH-AcOH) to give the sodium compound

Reaction (27 mg, mixture with

C (39 mg, 20%) as a colorless oil. MS (ESI - (M + H) ⁺ , (MH) ^- ).

1136.6 (2M + H)

MS (ESI): 565.5 (M-H) -.

567,, 4 1131., 8

D. (3S, 6R, 7S, 8S, 12R, 13S, 15S) -15-Amino-3,7-dihydroxy-12,13-epoxy-4,4,6,8 / 12,16-hexamethyl-17-acid - (2-hydroxymethyl-4-thiazolyl) -5-oxo-16 (E) -heptadecene

To a 10 mL round bottom flask charged with Compound C (40 mg, 71 mmol) and PtO ₂ (12 mg, 30% by weight) was added absolute ethanol (3 mL) under argon. The resulting black mixture was stirred under an atmosphere of hydrogen for 10 hours. The system was purged with nitrogen and the reaction mixture was filtered through a nylon membrane (washed with 25 mL of MeOH). The solvents were evaporated in vacuo to give compound D (29 mg, 76%) as a foam that was sufficiently pure to be used in the next step. LCMS: 541.3 (M + H) &lt; + & gt ^; .

E. [1S- [1R *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] - 7,11-dihydroxy-8,8,10,12,16-pentamethyl- 3- [1-methyl-2- (2-hydroxymethyl-4-thiazolyl) ethenyl] -4-aza-17-oxabicyclo [14.1.0] heptadecane-5,9-dione

A solution of compound D (29 mg, 54 mmol) in degassed DMF (21 mL) was treated with solid NaHCO ₃ (36 mg, 0.43 mmol) and diphenylphosphoryl azide (46 mL, 0.21 mmol) at 0 ° C under argon. . The resulting suspension was stirred at 4 ° C for 19 hours, cooled to -40 ° C, diluted with 25 mL of phosphate buffer pH 7 (carefully added to keep the internal temperature below -30 ° C) and extracted with EtOAc (4x10 ml). The organic extracts were washed with cold 10% aqueous LiCl solution (25 mL), dried (Na ₂ SO ₄ ) and evaporated in vacuo. The residue was purified using chromatotron (1 mm SiO ₂ , GF rotor, gradient elution with 2-5% MeOH-CHCl ₃ ) to give the title compound (9.1 mg, 34%) as a colorless oil. MS (ESI ⁺ ): 523.2 (M + H) ⁺ , MS (ESI -): 521.5 (MH) -.

Example 8

Me

[4S- [4R *, 7S *, 8R *, 9R *, 15R * (E)]] - 4,8-Dihydroxy-5,5,7,9,13-pentamethyl-16- [1-methyl-2] - (2-hydroxymethyl-4-thiazolyl) ethenyl] -1-aza-13 (Z) -cyclohexadecene-2,6-dione t-BuPh ₂ SiO b

HN.

Me

About OH O • ·

A. [1S- [1R *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] - 7,11-dihydroxy-8,8,10,12,16-pentamethyl- 3- [1-methyl-2- (2-tert-butyldiphenylsilyloxymethyl-4-thiazolyl) ethenyl] -4-aza-17-oxabicyclo [14.1.0] heptadecane-5,9- dion

A solution of 7E (6.8 mg, 13 mmol) in CH ₂ Cl ₂ (0.5 mL) was treated with triethylamine (2.7 mL, 20 mmol), 4-N, N- at 0 ° C under argon. dimethylaminopyridine (0.2 mg, 1.3 mmol); butyldiphenylsilyl chloride (3.7 mL, 14 mmol). The reaction mixture was gradually heated at 25 ° C (1 hour), cooled to 0 ° C, quenched with saturated aqueous NaHCO 3 (1 mL) and extracted with EtOAc (4x2 mL). The combined organic extracts were washed with brine (5 mL), dried (Na ₂ SO ₄ ) and evaporated in vacuo. The residue was purified by flash chromatography (SiO ₂ , 1.0x5 cm, gradient elution with 2 to 5% MeOH-CHCl ₃ ) to give compound A (7.0 mg, 71%) as a colorless oil. MS (ESI ⁺ ), 761.5 (M + H) ⁺ , MS (ESI), 759.7 (MH) -.

B. [4S- [4R *, 7S *, 8R *, 9R *, 15R * (E)]] - 4,8-Dihydroxy-5,5,7,9,13-pentamethyl-16- [1-methyl] -2- (2-hydroxymethyl-4-thiazolyl) ethenyl] -1-aza-13 (Z) -cyclohexadecene-2,6-dione

A solution of tungsten trichloride (0.10 g, 0.25 mmol) in anhydrous THF was treated with n-BuLi (1.6 M in hexane, 0.32 mL, 0.50 mmol) at -78 ° C under argon. The reaction mixture was heated at 25 ° C for 40 minutes and then re-cooled to 0 ° C. An aliquot (0.2 mL, 20 mmol) of the resulting dark green, homogeneous solution was added at 0 ° C under argon to a 3.55 cm ³ vial (1 dram) filled with Compound A (7.0 mg, 9.2 mmol). The reaction mixture was warmed to 25 ° C, stirred for 30 minutes, quenched with saturated aqueous NaHCO ₃ (0.5 mL) and extracted with EtOAc (4 x 1 mL). The combined organic extracts were dried (Na ₂ SO ₄ ) and evaporated in vacuo. The residue was purified by preparative TLC (SiO ₂ / 20x20x0.025 cm, eluting with 5% MeOH-CHCl ₃ ) to provide a divisible mixture of the silyl protected (13Z) isomer of Compound B and a small amount (<10%) of the minor (13E) isomer the protecting group was removed immediately in the next step.

A mixture of the silyl protected isomers of compound B (2.3 mg, 3.1 mmol) was treated with 0.3 mL of a buffered solution of HF-pyridine in THF (2: 1: 0.5 THF / pyridine / HF- solution at 25 ° C). pyridine from Aldrich Chemical Co.). After 1 h, the mixture was neutralized with saturated aqueous NaHCO 3 (0.5 mL) and extracted with EtOAc (4x1 mL). The combined organic extracts were washed with saturated aqueous NaHCO ₃ (1 mL), dried (Na ₂ SO ₄ ), and the volatiles were removed in vacuo. The residue was purified by preparative TLC (SiO ₂ , 20x10x0.025 cm, eluting with 5% MeOH-CHCl ₃ ) to give the title compound (13Z-isomer) along with an inseparable amount (<10%) of the minor (13E) isomer (0, 96 mg, 20% for two steps) as a thin film. MS (ESI ⁺ ): 507.3 (M + H) ^&lt; ^{+ &gt};, MS (ESI): 505.6 (MH).

Claims (6)

PATENT CLAIMS A compound of formula V wherein Q is selected from the group consisting of G is selected from the group consisting of alkyl, substituted alkyl, substituted or unsubstituted aryl, heterocyclo, RnPl2 Pl2 Ri Rn- i ^r R14 W is O or NR 6; X is 0 or Η, H; Y is selected from the group consisting of 0; H, OR 2, OR 2, 0R _1? ; NOR _1g ; H, NO ₁₉ ; H ₂ NR q R _21; H, H; or CHR ₂₂ ; OR ₁₇ OR ₁₇ may be a cyclic ketal; and Ζ ₂ are selected from the group consisting of CH ₂ , O, NR ₂₃ , S or SO ₂ , with only one skupina 1 or Z ₂ group being heteroatorial; _Β χ and B ₂ are selected from the group consisting of OR _24, or OCOR 5 ₂ or O ₂ CNR ₂ gR ₂₇ where B ₁ is hydrogen and Y is OH, H they can form a six-membered ring ketal or acetal; · D is selected from the group consisting of NR ₂ gR _2g, -NR q COR ^^ or saturated heterocycle; ^R l, ^R 2, ^R 3, ^R 4, ^R 5, ^R 6, ^R 7, ^R 13, ^R 14, ^R 18, ^R 19, ^R 2 0, ^r 21 ^r 22 ^r 26 ^{and R} 27 3 ^SOU selected from hydrogen, alkyl, substituted alkyl or aryl groups, and when R 1 and R _{2 are} alkyl may be joined to form cycloalkyl; when R ₃ and R _{4 are} alkyl, they may be joined to form cycloalkyl; Rg, R ₁₀ , R _1g , R _1? R _24, R ₂ 5 and R ₃₁ are selected from the group consisting of H, alkyl, substituted alkyl; R 8, ^R 11 ' ^R 12' ^R 28 ' ^R 30' ^R 32 ' ^R 33 ^{and R} 30 3 ^{SOU are} selected from the group consisting of H, alkyl, substituted alkyl, aryl, substituted aryl, cycloalkyl or heterocyclo; ^R 15 ^R 23 and ^R 29 3 ^are selected from the group consisting of Ή, alkyl, substituted alkyl, aryl, substituted aryl, cycloalkyl, heterocyclo, R ₃₂ C = O, R ₃ gSO ₂ , hydroxy, O-alkyl or O-substituted alkyl and pharmaceutically acceptable salts thereof, and any hydrates, solvates or geometric, optical and stereoisomers thereof With the proviso that compounds where W and X are both oxygen atoms; R 1, R 2 ^and R 7 are hydrogen atoms; R 8, R ₄ and R 8 are methyl; and R 8 is hydrogen or methyl; a and Z ₂ are CH ₂ ; and G is 1-methyl-2- (substituted-4-thiazolyl) ethenyl; Q has the same meaning as above. The compound of claim 1, wherein Q is X is an oxygen atom Y is an oxygen atom Z 1 and Z ₂ are CH ₂ and W is NR ₁₅ , 3. A compound selected from the group consisting of: [1S- [1R *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] -7.11 * dihydroxy-8,8,10,12, 16-Pentamethyl-3- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -4,13,17-trioxabicyclo [14.1.0] heptadecane-5,9-dione, [1S- [IR *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] -7,11-dihydroxy-8,8,10,12-tetramethyl-3- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -4,13,17-trioxabicyclo [14.1.0] heptadecane-5,9-dione, [4S- [4R *, 7S *, 8R *, 9R *, 15R * (E)]] - 4,8-Dihydroxy-5,5,7,9,13-pentamethyl-16- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -1,10 -dioxa-13-cyclohexadecene-2,6-dione, [4S- [4R *, 7S *, 8R *, 9R *, 15R * (E)]] - 4,8-dihydroxy-5,5,7,9 -tetramethyl-16- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -1,10-dioxa-13-cyclohexadecene-2,6-dione, [1S- [1R *, 3R * ( E), 7R *, 10S *, 11R *, 12R *, 16S *]] -7,11-dihydroxy-8,8,10,12,16-pentamethyl-3- [1-methyl-2- (2-methyl- 4-thiazolyl) ethenyl] -4,14,17-trioxabicyclo [14.1.0] heptadecane-5,9-dione, [1S- [1R *, 3R * (E), 7R *, 10S *, 11R *] 12R *, 16S *]] -7,11-dihydroxy-8,8,10,12-tetramethyl-3- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -4,14,17- trioxa bicyclo [14.1.0] heptadecane-5,9-dione, [4S- [4R *, 7S *, 8R *, 9R *, 15R * (E)]] - 4,8-dihydroxy-5,5, 7,9,13-Pentamethyl-16- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -1,11-dioxa-13-cyclohexadecene-2,6-dione, [4S- [4R] *, 7S *, 8R *, 9R *, 15R * (E)]] - 4,8-dihydroxy -5,5,7,9-tetramethyl-16- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -1,11-dioxa-13-cyclohexadecene-2,6-dione, [1S] - [IR *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] -7,11-dihydroxy-8,8,10,12,16-pentamethyl-3- [1-methyl -2- (2-methyl-4-thiazolyl) ethenyl] -4,17-dioxabicyclo [14.1.0] heptadecane-5,9-dione, [1S- [1R *, 3R * (E), 7R *] , 10S *, 11R *, 12R *, 16S *]] -7,11-dihydroxy-8,8,10,12-tetramethyl-3- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] - 4,17-dioxabicyclo [14.1.0] heptadecane-5,9-dione, [1S- [IR *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] - 7,11-dihydroxy-3,8,8,10,12,16-hexamethyl-3- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -4,17-dioxabicyclo [14,1,0 heptadecan-5,9-dione, [1S- [1R *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] -7,11-dihydroxy-3,8,8,10 12-pentamethyl-3- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -4,17-dioxabicyclo [14.1.0] heptadecane-5,9-dione, [4S- [ 4R *, 7S *, 8R *, 9R *, 15R * (E)]] - 4,8-dihydroxy-5,5,7,9,13,16-hexamethyl-16- [1-methyl-2- ( 2-methyl-4-thiazolyl) ethenyl] -1-oxa-13-cyclohexadecene-2,6-dione, [4S- [4R *, 7S *, 8R *, 9R *, 15R * (E)]] - 4 , 8-dihydr oxy-5,5,7,9,16-pentamethyl-16- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -1-oxa-13-cyclohexadecene-2,6-dione, [ 1S- [1R *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] - 7,11-dihydroxy-6,8,8,10,12,16-hexamethyl-3- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -4,17-dioxabicyclo [14.1.0] heptadecane-5,9-dione, [1S- [IR], 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] - 7,11-dihydroxy-6,8,8,10,12-pentamethyl-3- [1-methyl-2- (2-methyl)] -4-thiazolyl) ethenyl] -4,17-dioxabicyclo [14.1.0] heptadecane-5,9-dione, [4S- [4R *, 7S *, 8R *, 9R *, 15R * (E)] -4,8-Dihydroxy-5,5,7,9-tetramethyl-16- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -1-aza-13-cyclohexadecene-2,6 -dione, [1S- [1R *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] -7,11-dihydroxy-4,8,8,10,12,16-hexamethyl -3- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -4-aza-17-oxabicyclo [14.1.0] heptadecane-5,9-dione, [1S- [1R] , 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] - 7,11-dihydroxy-4,8,8,10,12-pentamethyl-3- [1-methyl-2- ( 2-methyl-4-thiazolyl) ethenyl] -4-aza-17-oxabicyclo [14.1.0] heptadecan-5,9-dione, [4S- [4R *, 7S *, 8R *, 9R *, 15R] * (E)]] - 4,8-dihydroxy- 1,5,5,7,9,13-hexamethyl-16- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -1-aza-13-cyclohexadecene-2,6-dione, [ 4S- [4R *, 7S *, 8R *, 9R *, 15R * (E)]] - 4,8-dihydroxy-1,5,5,7,9-pentamethyl-16- [1-methyl-2- (2-Methyl-4-thiazolyl) ethenyl] -1-aza-13-cyclohexadecene-2,6-dione, [4S- [4R *, 7S *, 8R *, 9R *, 15R * (E)]] - 4,8-dihydroxy-1,5,5,7,9-pentamethyl-16- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -1-aza-13-cyclohexadecene-2,6 -dione, [1S- [1R *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] -7,11-dihydroxy-8,8,10,12, 16-Pentamethyl-3- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -13-aza-4,17-dioxabicyclo [14.1.0] heptadecane-5,9-dione, [ 1S- [1R *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] -7,11-dihydroxy-8,8,10,12-tetramethyl-3- [1-methyl- 2- (2-methyl-4-thiazolyl) ethenyl] -13-aza-4,17-dioxabicyclo [14.1.0] heptadecane-5,9-dione, [4S- [4R *, 7S *, 8R *] , 9R *, 15R * (E)]] - 4,8-dihydroxy-5,5,7,9,13-pentamethyl-16- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -10-aza-1-oxa-13-cyclohexadecene-2,6-dione, [4S- [4R *, 7S *, 8R *, 9R *, 15R * (E)]] - 4,8-dihydroxy-5 5,7,9-tetramethyl-16- [1 methyl-2- (2-methyl-4-thiazolyl) ethenyl] -10-aza-1-oxa-13-cyclohexadecene-2,6-dione, [1S- [IR], 3R * (E), 7R * , 10S *, 11R *, 12R *, 16S *]] - 7,11-dihydroxy-8,8,10,12,16-pentamethyl-3- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl ] -14-aza-4,17-dioxabicyclo [14.1.0] heptadecane-5,9-dione, [1S- [1R *, 3R * (E), 7R *, 10S *, 11R *, 12R *] , 16S *]] -7,11-dihydroxy-8,8,10,12-tetramethyl-3- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -14-aza-4,17-dioxabicyclo [14,1,0] Heptadecane-5,9-dione, [4S- [4R *, 7S *, 8R *, 9R *, 15R * (E)]] - 4,8-dihydroxy-5,5,7 9,13-pentamethyl-16- [1-methyl-2- (2-methyl-4-thiazolyl) ethenyl] -1-aza-1-oxa-13-cyclohexadecene-2,6-dione; · T [4S- [4R *, 7S *, 8R *, 9R *, 15R * (E)]] - 4,8-dihydroxy-5,5,7,9-tetramethyl-16- [1-methyl- 2- (2-methyl-4-thiazolyl) ethenyl] -1-aza-1-oxa-13-cyclohexadecene-2,6-dione, [1S- [IR], 3R * (E), 7R *, 10S * 11R *, 12R *, 16S *]] - N-phenyl-7,11-dihydroxy-8,8,10,12,16-pentamethyl-5,9-dioxo4,17-dioxabicyclo [14.1.0] heptadecane- 3-carboxamide, [1S- [1R *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] - N-phenyl-7,11-dihydroxy-8,8,10,12- tetram ethyl-5,9-dioxo4,17-dioxabicyclo [14.1.0] heptadecane-3-carboxamide, [4S- [4R *, 7S *, 8R *, 9R *, 15R * (E)]] - N- phenyl-4,8-dihydroxy-5,5,7,9,13-pentamethyl-2,6-dioxo-1-oxa-13-cyclohexadecene-16-carboxamide, [4S- [4R *, 7S *, 8R *] , 9R *, 15R * (E)]] - N-phenyl-4,8-dihydroxy-5,5,7,9-tetramethyl-2,6-dioxo-1-oxa-13-cyclohexadecene-16-carboxamide, [1S- [1R *, 3R * (E), 7R *, 10S *, 11R *, 12R *, 16S *]] -7,11-dihydroxy-8, 8,10,12,16-pentamethyl-3- [1S-] methyl-2- (2-methyl-4-thiazolyl) cyclopropyl] -4,17-dioxabicyclo [14.1.0] heptadecane-5,9-dione, [1S- [IR], 3R * (E) , 7R *, 10S *, 11R *, 12R *, 16S *]] -7,1-dihydroxy-8, 8,10,12-tetramethyl-3- [1-methyl-2- (2-methyl-4-thiazolyl)] cyclopropyl] -4,17-dioxabicyclo [14.1.0] heptadecane-5,9-dione, [4S [4R *, 7S *, 8R *, 9R *, 15R * (E)]] - 4,8- dihydroxy-5,5,7,9,13-pentamethyl-16- [1-methyl-2- (hydroxymethyl-4-thiazolyl) ethenyl] -1-aza-13 (Z) -cyclohexadecene-2,6-dione And pharmaceutically acceptable salts, solvates and hydrates thereof. 4. A method of treating cancer in a patient comprising administering to said patient an effective amount of a compound of claim 1. 5. A method of treating a hyperproliferative disorder in a patient comprising administering to said patient an effective amount of a compound of claim 1. A method of providing an anti-angiogenic effect in a patient in need of such treatment, comprising administering to said patient an effective amount of a compound of claim 1.